Title:
SIGNAL-SENSOR POLYNUCLEOTIDES FOR THE ALTERATION OF CELLULAR PHENOTYPES
Kind Code:
A1


Abstract:
The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of signal-sensor polynucleotides, primary transcripts and mmRNA molecules.



Inventors:
Hoge, Stephen G. (New York, NY, US)
Chakraborty, Tirtha (Medford, MA, US)
Frederick, Joshua P. (Boston, MA, US)
John, Matthias (Cambridge, MA, US)
De Fougerolles, Antonin (Brookline, MA, US)
Application Number:
14/041011
Publication Date:
07/17/2014
Filing Date:
09/30/2013
Assignee:
MODERNA THERAPEUTICS, INC. (Cambridge, MA, US)
Primary Class:
Other Classes:
435/320.1
International Classes:
A61K48/00; C12N15/85
View Patent Images:



Other References:
Smirnov et al. (The American Journal of Human Genetics, 2008, 83:243-253)
NCBI (2014, Reference Sequence: NM_003326.4)
Suzuki et al. (Molecular Therapy 2008, 16:1719-1726)
Kron et al. (Molecular Therapy, 2011, 19:1547-1557)
Andarini (Cancer Research, 2004, 64:3281-3287)
Rotondaro et al. (Gene, 1996, 168:195-198)
Suzuki et al. (Molecular Therapy, 2008, 16:1719-1726)
Singh et al. (ACS Nano, 2008, 2:1040-1050).
Primary Examiner:
WU, JULIE ZHEN QIN
Attorney, Agent or Firm:
Cooley LLP/ModernaTX, Inc. (Washington, DC, US)
Claims:
We claim:

1. An isolated synthetic signal-sensor polynucleotide, wherein said isolated synthetic signal-sensor polynucleotide comprises an mRNA which encodes an oncology-related polypeptide of interest and one or more sensor sequences selected from the group consisting of any of SEQ ID NOs: 3529-4549, SEQ ID NOs: 5571-6591 and functional variants thereof.

2. The isolated synthetic signal-sensor polynucleotide of claim 1 wherein the oncology-related polypeptide of interest is selected from the group consisting of SEQ ID NOs: 1321-2487, 6611-6616, 7355-7361, 7490, 7492, 7493, 7512, 7514, 7516 and 7517.

3. The isolated synthetic signal-sensor polynucleotide of claim 1 wherein the mRNA comprises at least an open reading frame of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2488-2496, 6617-6621, 7348-7354, 7362-7489, 7491, 7494, 7506, 7511 and 7513.

4. The isolated synthetic signal-sensor polynucleotide of claim 3, wherein the open reading frame is codon optimized.

5. The isolated synthetic signal-sensor polynucleotide of claim 1, wherein the mRNA comprises two stop codons.

6. The isolated synthetic signal-sensor isolated polynucleotide of claim 1, wherein the mRNA comprises a first stop codon “TGA” and a second stop codon selected from the group consisting of “TAA,” “TGA” and “TAG.”

7. The isolated synthetic signal-sensor polynucleotide of claim 1, wherein the mRNA has a 3′ tailing sequence of linked nucleosides selected from the group consisting of a poly-A tail of at least 140 nucleotides, a triple helix, and a poly A-G quartet.

8. The isolated synthetic signal-sensor polynucleotide of claim 1, wherein the mRNA comprises at least one 5′terminal cap selected from the group consisting of Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

9. The isolated synthetic signal-sensor polynucleotide of claim 1, where the isolated synthetic signal-sensor polynucleotide is substantially purified.

10. The isolated synthetic signal-sensor polynucleotide of claim 1, wherein the isolated synthetic signal-sensor polynucleotide comprises at least one chemical modification.

11. The isolated synthetic signal-sensor polynucleotide of claim 10, wherein the at least one chemical modification is 1-methylpseudouridine.

12. The isolated synthetic signal-sensor polynucleotide of claim 11, further comprising the chemical modification 5-methylcytidine.

13. The isolated synthetic signal-sensor polynucleotide of claim 1, where the isolated synthetic signal-sensor polynucleotide comprises at least two chemical modifications.

14. The isolated synthetic signal-sensor polynucleotide of claim 13, wherein the modifications are located on one or more of a nucleoside and/or the backbone of said nucleotides.

15. The isolated synthetic signal-sensor polynucleotide of claim 13, where the modifications are located on both a nucleoside and a backbone linkage.

16. The isolated synthetic signal-sensor polynucleotide of claim 13, where the modifications are located on the backbone linkage.

17. The isolated synthetic signal-sensor polynucleotide of claim 1, wherein the isolated signal-sensor polynucleotide is codon optimized.

18. The isolated synthetic signal-sensor polynucleotide of claim 1, wherein the isolated signal-sensor polynucleotide is formulated.

19. The isolated synthetic signal-sensor polynucleotide of claim 1 wherein the polypeptide of interest is a factor modulating the affinity between HIF subunits and/or HIF-dependent gene expression.

20. The isolated synthetic signal-sensor polynucleotide of claim 19 wherein the HIF subunits are selected from the group consisting of SEQ ID NO: 6611-6616.

21. The isolated synthetic signal-sensor polynucleotide of claim 1 wherein the isolated synthetic signal-sensor polynucleotide comprises at least one translation enhancer element.

22. An isolated synthetic signal-sensor polynucleotide comprising: (a) a first region of linked nucleosides, said first region encoding an oncology-related polypeptide of interest selected from the group consisting of SEQ ID NOs: 1321-2487, 6611-6616 and 7355-7361, 7490, 7492, 7493, 7512, 7514, 7516 and 7517; (b) a first flanking region located 5′ relative to said first region comprising; (i) a sequence of linked nucleosides selected from the group consisting of the native 5′ untranslated region (UTR) of any of the nucleic acids that encode any of SEQ ID NOs: 1321-2487, 6611-6616, 7355-7361, 7490, 7492, 7493, 7512, 7514, 7516, 7517, SEQ ID NO: 1-4 and functional variants thereof; (c) a second flanking region located 3′ relative to said first region comprising; (i′) a sequence of linked nucleosides selected from the group consisting of the native 3′ UTR of any of the nucleic acids that encode any of SEQ ID NOs: 1321-2487, 6611-6616, 7355-7361, 7490, 7492, 7493, 7512, 7514, 7516, 7517, SEQ ID NO: 5-21 and functional variants thereof; (ii′) one or more sensor sequences located selected from the group consisting of the any of SEQ ID NOs: 3529-4549, SEQ ID NOs: 5571-6591 and functional variants thereof; and (iii′) a 3′ tailing sequence of linked nucleosides.

23. The isolated synthetic signal-sensor polynucleotide of claim 22 wherein the first region of linked nucleosides comprises at least an open reading frame of a nucleic acid sequence, wherein the nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 2488-2496, 6617-6621, 7348-7354, 7362-7489, 7491, 7494, 7506, 7511 and 7513.

24. The isolated synthetic signal-sensor polynucleotide of claim 23, wherein the open reading frame is codon optimized.

25. The isolated synthetic signal-sensor polynucleotide of claim 22, wherein the first region comprises two stop codons.

26. The isolated synthetic signal-sensor isolated polynucleotide of claim 22, wherein the first region comprises a first stop codon “TGA” and a second stop codon selected from the group consisting of “TAA,” “TGA” and “TAG.”

27. The isolated synthetic signal-sensor polynucleotide of claim 22, wherein the 3′ tailing sequence of linked nucleosides is selected from the group consisting of a poly-A tail of at least 140 nucleotides, a triple helix, and a poly A-G quartet.

28. The isolated synthetic signal-sensor polynucleotide of claim 22, wherein the first flanking region further comprises at least one 5′terminal cap.

29. The isolated synthetic signal-sensor polynucleotide of claim 28, wherein the at least one 5′ terminal cap is selected from the group consisting of Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

30. The isolated synthetic signal-sensor polynucleotide of claim 28 where the isolated signal-sensor polynucleotide is substantially purified.

31. The isolated synthetic signal-sensor polynucleotide of claim 22, wherein the isolated synthetic signal-sensor polynucleotide comprises at least one chemical modification.

32. The isolated synthetic signal-sensor polynucleotide of claim 31, wherein the at least one chemical modification is 1-methylpseudouridine.

33. The isolated synthetic signal-sensor polynucleotide of claim 32, further comprising the chemical modification 5-methylcytidine.

34. The isolated synthetic signal-sensor polynucleotide of claim 22, comprising at least two chemical modifications in the first region.

35. The isolated synthetic signal-sensor polynucleotide of claim 34, wherein the modifications are located on one or more of a nucleoside and/or the backbone of said nucleotides.

36. The isolated synthetic signal-sensor polynucleotide of claim 34, where the modifications are located on both a nucleoside and a backbone linkage.

37. The isolated synthetic signal-sensor polynucleotide of claim 34, where the modifications are located on the backbone linkage.

38. The isolated synthetic signal-sensor polynucleotide of claim 22, where the isolated signal-sensor polynucleotide is codon optimized.

39. The isolated synthetic signal-sensor polynucleotide of claim 38, wherein the first region of linked nucleosides is codon optimized.

40. The isolated synthetic signal-sensor polynucleotide of claim 22, wherein the isolated signal-sensor polynucleotide is formulated.

41. A method of treating a disease, disorder and/or condition in a subject in need thereof by increasing the level of an oncology-related polypeptide of interest comprising administering to said subject an isolated synthetic signal-sensor polynucleotide encoding said oncology-related polypeptide.

42. A method of reducing, eliminating or preventing tumor growth in a subject in need thereof by increasing the level of an oncology-related polypeptide of interest comprising administering to said subject an isolated synthetic signal-sensor polynucleotide encoding said oncology-related polypeptide.

43. A method of reducing and/or ameliorating at least one symptom of cancer in a subject need thereof by increasing the level of an oncology-related polypeptide of interest comprising administering to said subject an isolated synthetic signal-sensor polynucleotide encoding said oncology-related polypeptide.

44. The method of any of claims 41-43 wherein the disease, disorder and/or condition is selected from the group consisting of adrenal cortical cancer, advanced cancer, anal cancer, aplastic anemia, bileduct cancer, bladder cancer, bone cancer, bone metastasis, brain tumors, brain cancer, breast cancer, childhood cancer, cancer of unknown primary origin, Castleman disease, cervical cancer, colon/rectal cancer, endometrial cancer, esophagus cancer, Ewing family of tumors, eye cancer, gallbladder cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gestational trophoblastic disease, Hodgkin disease, Kaposi sarcoma, renal cell carcinoma, laryngeal and hypopharyngeal cancer, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myelomonocytic leukemia, liver cancer, hepatocellular carcinoma (HCC), non-small cell lung cancer, small cell lung cancer, lung carcinoid tumor, lymphoma of the skin, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, oral cavity and oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumors, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma in adult soft tissue, basal and squamous cell skin cancer, melanoma, small intestine cancer, stomach cancer, testicular cancer, throat cancer, thymus cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, Wilms tumor and secondary cancers caused by cancer treatment.

45. The method of claim 44 wherein the tumor growth is results from a disease, disorder and/or condition selected from the group consisting of adrenal cortical cancer, advanced cancer, anal cancer, aplastic anemia, bileduct cancer, bladder cancer, bone cancer, bone metastasis, brain tumors, brain cancer, breast cancer, childhood cancer, cancer of unknown primary origin, Castleman disease, cervical cancer, colon/rectal cancer, endometrial cancer, esophagus cancer, Ewing family of tumors, eye cancer, gallbladder cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gestational trophoblastic disease, Hodgkin disease, Kaposi sarcoma, renal cell carcinoma, laryngeal and hypopharyngeal cancer, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myelomonocytic leukemia, liver cancer, hepatocellular carcinoma (HCC), non-small cell lung cancer, small cell lung cancer, lung carcinoid tumor, lymphoma of the skin, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, oral cavity and oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumors, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma in adult soft tissue, basal and squamous cell skin cancer, melanoma, small intestine cancer, stomach cancer, testicular cancer, throat cancer, thymus cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, Wilms tumor and secondary cancers caused by cancer treatment.

46. The method of any of claims 41-43 wherein the administration of the isolated synthetic signal-sensor polynucleotide reduces the number of cancer cells, eliminates cancer cells, prevents an increase in cancer cells and/or alleviates the symptoms of cancer in a subject.

47. The method of claim 43 wherein the at least one symptom of cancer is selected from the group consisting of weakness, aches and pains, fever, fatigue, weight loss, blood clots, increased blood calcium levels, low white blood cell count, short of breath, dizziness, headaches, hyperpigmentation, jaundice, erthema, pruritis, excessive hair growth, change in bowel habits, change in bladder function, long-lasting sores, white patches inside the mouth, white spots on the tongue, unusual bleeding or discharge, thickening or lump on parts of the body, indigestion, trouble swallowing, changes in warts or moles, change in new skin and nagging cough and hoarseness.

48. The methods of any of claims 41-43, wherein the isolated synthetic signal-sensor polynucleotide is formulated.

49. The method of claim 48, wherein the isolated synthetic signal-sensor polynucleotide is administered at a total daily dose of between 0.001 ug and 150 ug.

50. The method of claim 49, wherein administration is by injection, topical administration, ophthalmic administration or intranasal administration.

51. The method of claim 50, wherein administration is by injection and said injection is selected from the group consisting of intradermal, subcutaneous and intramuscular.

52. The method of claim 50, wherein administration is topical administration and said topical administration is selected from the group consisting of cream, lotion, ointment, gel, spray, solution and the like.

53. A method of preferentially inducing cell death in cancer cells in a tissue or organ, comprising (a) contacting said tissue or organ with an isolated synthetic signal-sensor polynucleotide, wherein said isolated synthetic signal-sensor polynucleotide encodes (i) an oncology-related polypeptide whose expression triggers apoptosis or cell death, and (ii) at least one microRNA binding site of a microRNA, where the expression of said microRNA in the cancer cell is lower than the expression of said microRNA in normal, non cancerous cells.

Description:

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No. 61/753,661, filed Jan. 17, 2013, entitled Signal-Sensor Polynucleotides for the Alternation of Cellular Phenotypes and Microenvironments; U.S. Provisional Application No. 61/754,159, filed Jan. 18, 2013, entitled Signal-Sensor Polynucleotides for the Alternation of Cellular Phenotypes and Microenvironments; U.S. Provisional Application No. 61/781,097, filed Mar. 14, 2013, entitled Signal-Sensor Polynucleotides for the Alternation of Cellular Phenotypes and Microenvironments; U.S. Provisional Application No. 61/829,334, filed May 31, 2013, entitled Signal-Sensor Polynucleotides for the Alternation of Cellular Phenotypes and Microenvironments; U.S. Provisional Application No. 61/839,893, filed Jun. 27, 2013, entitled Signal-Sensor Polynucleotides for the Alternation of Cellular Phenotypes and Microenvironments; U.S. Provisional Application No. 61/842,733, filed Jul. 3, 2013, entitled Signal-Sensor Polynucleotides for the Alternation of Cellular Phenotypes and Microenvironments; and U.S. Provisional Application No. 61/857,304, filed Jul. 23, 2013, entitled Signal-Sensor Polynucleotides for the Alternation of Cellular Phenotypes and Microenvironments; the contents of each of which is herein incorporated by reference in its entirety.

REFERENCE TO SEQUENCE LISTING

The present application is being filed along with a Sequence Listing in electronic format. The Sequence listing file, entitled M37US.txt, was created on Sep. 30, 2013 and is 9,748,568 bytes in size. The information in electronic format of the Sequence Listing is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

The invention relates to compositions, methods, processes, kits and devices for the design, preparation, manufacture and/or formulation of signal-sensor polynucleotides, primary constructs and mRNA molecules for the alteration of cellular phenotypes and micro environments.

BACKGROUND OF THE INVENTION

Cancer is a disease characterized by uncontrolled cell division and growth within the body. In the United States, roughly a third of all women and half of all men will experience cancer in their lifetime. Polypeptides are involved in every aspect of the disease including cancer cell biology (carcinogenesis, cell cycle suppression, DNA repair and angiogenesis), treatment (immunotherapy, hormone manipulation, enzymatic inhibition), diagnosis and determination of cancer type (molecular markers for breast, prostate, colon and cervical cancer for example). With the host of undesired consequences brought about by standard treatments such as chemotherapy and radiotherapy used today, genetic therapy for the manipulation of disease-related peptides and their functions provides a more targeted approach to disease diagnosis, treatment and management.

To this end, it has been previously shown that certain modified mRNA sequences have the potential as therapeutics with benefits beyond just evading, avoiding or diminishing the immune response. Such studies are detailed in published co-pending International Publication No WO2012019168 filed August 5, 201, International Publication No WO2012045082 filed Oct. 3, 2011, International Publication No WO2012045075 filed Oct. 3, 2011, International Publication No WO2013052523 filed Oct. 3, 2012, and International Publication No WO2013090648 filed Dec. 14, 2012 the contents of which are incorporated herein by reference in their entirety.

The use of modified polynucleotides in the fields of antibodies, viruses, veterinary applications and a variety of in vivo settings have been explored and are disclosed in, for example, co-pending and co-owned U.S. Provisional Patent Application No. 61/618,862, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Biologics; U.S. Provisional Patent Application No. 61/681,645, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Biologics; U.S. Provisional Patent Application No. 61/737,130, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Biologics; U.S. Provisional Patent Application No. 61/618,866, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Antibodies; U.S. Provisional Patent Application No. 61/681,647, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Antibodies; U.S. Provisional Patent Application No. 61/737,134, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Antibodies; U.S. Provisional Patent Application No. 61/618,868, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Vaccines; U.S. Provisional Patent Application No. 61/681,648, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Vaccines; U.S. Provisional Patent Application No. 61/737,135, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Vaccines; U.S. Provisional Patent Application No. 61/618,870, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Therapeutic Proteins and Peptides; U.S. Provisional Patent Application No. 61/681,649, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Therapeutic Proteins and Peptides; U.S. Provisional Patent Application No. 61/737,139, filed Dec. 14, 2012, Modified Polynucleotides for the Production of Therapeutic Proteins and Peptides; U.S. Provisional Patent Application No. 61/618,873, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Secreted Proteins; U.S. Provisional Patent Application No. 61/681,650, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Secreted Proteins; U.S. Provisional Patent Application No. 61/737,147, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Secreted Proteins; U.S. Provisional Patent Application No. 61/618,878, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Plasma Membrane Proteins; U.S. Provisional Patent Application No. 61/681,654, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Plasma Membrane Proteins; U.S. Provisional Patent Application No. 61/737,152, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Plasma Membrane Proteins; U.S. Provisional Patent Application No. 61/618,885, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Cytoplasmic and Cytoskeletal Proteins; U.S. Provisional Patent Application No. 61/681,658, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Cytoplasmic and Cytoskeletal Proteins; U.S. Provisional Patent Application No. 61/737,155, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Cytoplasmic and Cytoskeletal Proteins; U.S. Provisional Patent Application No. 61/618,896, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Intracellular Membrane Bound Proteins; U.S. Provisional Patent Application No. 61/668,157, filed Jul. 5, 2012, entitled Modified Polynucleotides for the Production of Intracellular Membrane Bound Proteins; U.S. Provisional Patent Application No. 61/681,661, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Intracellular Membrane Bound Proteins; U.S. Provisional Patent Application No. 61/737,160, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Intracellular Membrane Bound Proteins; U.S. Provisional Patent Application No. 61/618,911, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Nuclear Proteins; U.S. Provisional Patent Application No. 61/681,667, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Nuclear Proteins; U.S. Provisional Patent Application No. 61/737,168, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Nuclear Proteins; U.S. Provisional Patent Application No. 61/618,922, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Proteins; U.S. Provisional Patent Application No. 61/681,675, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Proteins; U.S. Provisional Patent Application No. 61/737,174, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Proteins; U.S. Provisional Patent Application No. 61/618,935, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. Provisional Patent Application No. 61/681,687, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. Provisional Patent Application No. 61/737,184, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. Provisional Patent Application No. 61/618,945, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. Provisional Patent Application No. 61/681,696, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. Provisional Patent Application No. 61/737,191, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. Provisional Patent Application No. 61/618,953, filed Apr. 2, 2012, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. Provisional Patent Application No. 61/681,704, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. Provisional Patent Application No. 61/737,203, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. Provisional Patent Application No. 61/681,720, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Cosmetic Proteins and Peptides; U.S. Provisional Patent Application No. 61/737,213, filed Dec. 14, 2012, entitled Modified Polynucleotides for the Production of Cosmetic Proteins and Peptides; U.S. Provisional Patent Application No. 61/681,742, filed Aug. 10, 2012, entitled Modified Polynucleotides for the Production of Oncology-Related Proteins and Peptides; International Application No PCT/US2013/030062, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Biologics and Proteins Associated with Human Disease; U.S. patent application Ser. No. 13/791,922, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Biologics and Proteins Associated with Human Disease; International Application No PCT/US2013/030063, filed Mar. 9, 2013, entitled Modified Polynucleotides; International Application No. PCT/US2013/030064, entitled Modified Polynucleotides for the Production of Secreted Proteins; U.S. patent application Ser. No. 13/791,921, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Secreted Proteins; International Application No PCT/US2013/030059, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Membrane Proteins; International Application No. PCT/US2013/030066, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Cytoplasmic and Cytoskeletal Proteins; International Application No. PCT/US2013/030067, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Nuclear Proteins; International Application No. PCT/US2013/030060, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Proteins; International Application No. PCT/US2013/030061, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; U.S. patent application Ser. No. 13/791,910, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Proteins Associated with Human Disease; International Application No. PCT/US2013/030068, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Cosmetic Proteins and Peptides; and International Application No. PCT/US2013/030070, filed Mar. 9, 2013, entitled Modified Polynucleotides for the Production of Oncology-Related Proteins and Peptides; International Patent Application No. PCT/US2013/031821, filed Mar. 15, 2013, entitled In Vivo Production of Proteins; the contents of each of which are herein incorporated by reference in their entireties.

Formulations and delivery of modified polynucleotides are described in, for example, co-pending and co-owned International Publication No WO2013090648, filed Dec. 14, 2012, entitled Modified Nucleoside, Nucleotide, Nucleic Acid Compositions and US Publication No US20130156849, filed Dec. 14, 2012, entitled Modified Nucleoside, Nucleotide, Nucleic Acid Compositions; the contents of each of which are herein incorporated by reference in their entireties.

The next generation of therapeutics must also address the complex cellular microenvironment of the cancer and have the capacity for cell, tissue, organ or patient stratification, whether structurally or functionally.

The present invention addresses this need by providing nucleic acid based compounds or polynucleotide-encoding nucleic acid-based compounds (e.g., signal-sensor polynucleotides) which encode a polypeptide of interest and which have structural and/or chemical features that allow for greater selectivity, profiling or stratification along defineable disease characteristics or metrics.

SUMMARY OF THE INVENTION

Described herein are compositions, methods, processes, kits and devices for the design, preparation, manufacture and/or formulation of signal-sensor polynucleotide molecules encoding at least one oncology-related polypeptide of interest. Such signal-sensor polynucleotides may be chemically modified mRNA (mmRNA) molecules.

The present invention provides an isolated signal-sensor polynucleotide comprising a region encoding an oncology-related polypeptide of interest that functions, when translated, to send a death or survival signal. Such death or survival signals include those which (i) alter (increase or decrease) the expression of one or more proteins, nucleic acids, or non-coding nucleic acids, (ii) alter the binding properties of biomolecules within the cell, and/or (iii) perturb the cellular microenvironment in a therapeutically beneficial way.

Optionally, the signal-sensor polynucleotide may also encode in a flanking region, one or more sensor sequences. Such sensor sequences function to “sense” the cell, tissue or organ microenvironment and confer upon the signal-sensor polynucleotide an altered expression or half life profile (increased or decreased) depending on the interactions of the sensor sequence with the cell, tissue or organ microenvironment.

In one aspect, provided herein are signal-sensor polynucleotide comprising, a first region of linked nucleosides, a first flanking region located 5′ relative to said first region and a second flanking region located 3′ relative to said first region. The first region may encode an oncology-related polypeptide of interest such as, but not limited to, SEQ ID NOs: 1321-2487, 6611-6616 and 7355-7361, 7490, 7492, 7493, 7512, 7514, 7516 and 7517 and the first flanking region may include a sequence of linked nucleosides such as, but not limited to, the native 5′ untranslated region (UTR) of any of the nucleic acids that encode any of SEQ ID NOs: 1321-2487, 6611-6616, 7355-7361, 7490, 7492, 7493, 7512, 7514, 7516, 7517, SEQ ID NO: 1-4 and functional variants thereof. The first region may comprise at least an open reading frame of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2488-2496, 6617-6621, 7348-7354, 7362-7489, 7491, 7494, 7506, 7511 and 7513.

The second flanking region may include a sequence of linked nucleosides such as, but not limited to, the native 3′ UTR of any of the nucleic acids that encode any of SEQ ID NOs: 1321-2487, 6611-6616, 7355-7361, 7490, 7492, 7493, 7512, 7514, 7516, 7517, SEQ ID NO: 5-21 and functional variants thereof, and one or more sensor sequences located such as, but not limited to, SEQ ID NOs: 3529-4549, SEQ ID NOs: 5571-6591 and functional variants thereof. The signal-sensor polynucleotide may also include a 3′ tailing sequence of linked nucleosides.

In another aspect, provided herein is a signal-sensor polynucleotide which comprises an mRNA encoding an oncology-related polypeptide of interest and one or more sensor sequences such as, but not limited to, SEQ ID NOs: 3529-4549, SEQ ID NOs: 5571-6591 and functional variants thereof. The oncology-related polypeptide of interest may be, but is not limited to, SEQ ID NOs: 1321-2487, 6611-6616, 7355-7361, 7490, 7492, 7493, 7512, 7514, 7516 and 7517. The mRNA may include at least one open reading frame of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2488-2496, 6617-6621, 7348-7354, 7362-7489, 7491, 7494, 7506, 7511 and 7513.

The signal-sensor polynucleotides may comprise one, two, three or more than three stop codons. In one aspect, the signal-sensor polynucleotides comprise two stop codons. As a non-limiting example, the first stop codon is “TGA” and the second stop codon is selected from the group consisting of “TAA,” “TGA” and “TAG.” In another aspect, signal-sensor polynucleotides comprise three stop codons.

The signal-sensor polynucleotides may have a 3′ tailing sequence of linked nucleosides such as, but not limited to, a poly-A tail of at least 140 nucleotides, a triple helix, and a poly A-G quartet.

The signal-sensor polynucleotides may have a 5′cap such as, but not limited to, Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

In one aspect, the signal-sensor polynucleotides may include at least one chemical modification such as, but not limited to, modifications located on one or more of a nucleoside and/or the backbone of the nucleotides. In one embodiment, the signal-sensor polynucleotides comprise a pseudouridine analog such as, but not limited to, 1-carboxymethyl-pseudouridine, 1-propynyl-pseudouridine, 1-taurinomethyl-pseudouridine, 1-taurinomethyl-4-thio-pseudouridine, 1-methyl-pseudouridine (m1ψ), 1-methyl-4-thio-pseudouridine (m1s4ψ), 4-thio-1-methyl-pseudouridine, 3-methyl-pseudouridine (m3ψ), 2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydropseudouridine, 2-thio-dihydropseudouridine, 2-methoxyuridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, N1-methyl-pseudouridine, 1-methyl-3-(3-amino-3-carboxypropyl)pseudouridine (acp3ψ), and 2′-O-methyl-pseudouridine (ψm). In another embodiment, the signal-sensor polynucleotides comprise the pseudouridine analog 1-methylpseudouridine. In yet another embodiment, the signal-sensor polynucleotides comprise the pseudouridine analog 1-methylpseudouridine and the modified nucleoside 5-methylcytidine.

In another aspect, the signal-sensorpolynucleotides may include at least two chemical modifications such as, but not limited to, modifications located on one or more of a nucleoside and/or the backbone of the nucleotides. As a non-limiting example, the signal-sensor polynucleotide comprises the chemical modifications 1-methylpseudouridine and 5-methylcytidine.

The signal-sensor polynucleotides may comprise at least one translation enhancer element (TEE) such as, but not limited to, TEE-001-TEE-705.

In one aspect, the signal-sensor polynucleotide encodes a factor modulating the affinity between HIF subunits and/or HIF-dependent gene expression such as, but not limited to, SEQ ID NO: 6611-6616.

The signal-sensor polynucleotides may be purified and/or formulated.

Employing the signal-sensor polynucleotides, the present invention provides a method of treating a disease, disorder and/or condition in a subject in need thereof by increasing the level of an oncology-related polypeptide of interest comprising administering to said subject an isolated signal-sensor polynucleotide encoding said oncology-related polypeptide. The disease, disorder and/or condition may include, but is not limited to, adrenal cortical cancer, advanced cancer, anal cancer, aplastic anemia, bileduct cancer, bladder cancer, bone cancer, bone metastasis, brain tumors, brain cancer, breast cancer, childhood cancer, cancer of unknown primary origin, Castleman disease, cervical cancer, colon/rectal cancer, endometrial cancer, esophagus cancer, Ewing family of tumors, eye cancer, gallbladder cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gestational trophoblastic disease, Hodgkin disease, Kaposi sarcoma, renal cell carcinoma, laryngeal and hypopharyngeal cancer, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myelomonocytic leukemia, liver cancer, non-small cell lung cancer, small cell lung cancer, lung carcinoid tumor, lymphoma of the skin, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, oral cavity and oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumors, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma in adult soft tissue, basal and squamous cell skin cancer, melanoma, small intestine cancer, stomach cancer, testicular cancer, throat cancer, thymus cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, Wilms tumor and secondary cancers caused by cancer treatment.

The present invention provides a method of reducing, eliminating, or preventing tumor growth in a subject in need thereof by increasing the level of an oncology-related polypeptide of interest comprising administering to said subject an isolated signal-sensor polynucleotide encoding said oncology-related polypeptide. The tumor growth may be associated with or results from a disease, disorder and/or condition such as, but not limited to, adrenal cortical cancer, advanced cancer, anal cancer, aplastic anemia, bileduct cancer, bladder cancer, bone cancer, bone metastasis, brain tumors, brain cancer, breast cancer, childhood cancer, cancer of unknown primary origin, Castleman disease, cervical cancer, colon/rectal cancer, endometrial cancer, esophagus cancer, Ewing family of tumors, eye cancer, gallbladder cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gestational trophoblastic disease, Hodgkin disease, Kaposi sarcoma, renal cell carcinoma, laryngeal and hypopharyngeal cancer, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myelomonocytic leukemia, liver cancer, non-small cell lung cancer, small cell lung cancer, lung carcinoid tumor, lymphoma of the skin, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, oral cavity and oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumors, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma in adult soft tissue, basal and squamous cell skin cancer, melanoma, small intestine cancer, stomach cancer, testicular cancer, throat cancer, thymus cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, Wilms tumor and secondary cancers caused by cancer treatment.

The present invention provides a method of reducing and/or ameriorating at least one symptom of cancer in a subject in need thereof by increasing the level of a polypeptide of interest comprising administering to said subject an isolated signal-sensor polynucleotide encoding said oncology-related polypeptide. Non-limiting examples of symptoms include weakness, aches and pains, fever, fatigue, weight loss, blood clots, increased blood calcium levels, low white blood cell count, short of breath, dizziness, headaches, hyperpigmentation, jaundice, erthema, pruritis, excessive hair growth, change in bowel habits, change in bladder function, long-lasting sores, white patches inside the mouth, white spots on the tongue, unusual bleeding or discharge, thickening or lump on parts of the body, indigestion, trouble swallowing, changes in warts or moles, change in new skin and nagging cough and hoarseness.

The present invention provides a method of preferentially inducing cell death in cancer cells in a tissue or organ comprising contacting the tissue or organ with a signal-sensor polynucleotide encoding an oncology-related polypeptide whose expression triggers apoptosis or cell death and at least one microRNA binding site of a microRNA where the expression of the microRNA in the cancer cell is lower than the expression of the mircroRNA in normal non-cancerous cells.

The signal-sensor polynucleotide may be administered at a total daily dose of between 0.001 ug and 150 ug. Administration of a signal-sensor polynucleotide may be by injection, topical administration, ophthalmic administration or intranasal administration. In one aspect, administration may be by injection such as, but not limited to, intradermal, subcutaneous and intramuscular. In another aspect, administration may be topical such as, but not limited to, using creams, lotions, ointments, gels, sprays, solutions and the like.

The details of various embodiments of the invention are set forth in the description below. Other features, objects, and advantages of the invention will be apparent from the description and the drawings, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing and other objects, features and advantages will be apparent from the following description of particular embodiments of the invention, as illustrated in the accompanying drawings in which like reference characters refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of various embodiments of the invention.

FIG. 1 is a schematic of a primary construct of the present invention.

FIG. 2 is an expanded schematic of the second flanking region of a primary construct of the present invention illustrating the signal-sensor elements of the polynucleotide.

FIG. 3 is a gel profile of Apoptosis-Inducing Factor short (AIFsh) protein from AIFsh modified mRNA in mammals. FIG. 3A shows the expected size of AIFsh. FIG. 3B shows the expected size of AIFsh.

FIG. 4 is a gel profile of Siah E3 ubiquitin protein ligase 1 (SIAH1) protein from SIAH1 modified mRNA in mammals. FIG. 4A shows the expected size of SIAH1. FIG. 4B shows the expected size of SIAH1.

FIG. 5 is a gel profile of constitutively active (C.A.) caspase 3 (also known as reverse caspase 3 (Rev-Caspase 3)) protein from C.A. caspase 3 modified mRNA in mammals. FIG. 5A shows the expected size of C.A. caspase 3. FIG. 5B shows the expected size of C.A. caspase 3.

FIG. 6 is a gel profile of Granulysin protein from granulysin modified mRNA in mammals. FIG. 6A shows the expected size of granulysin. FIG. 6B shows the expected size of granulysin.

FIG. 7 is a western blot of C.A. caspase 3 and C.A. caspase 6. FIG. 7A shows protein from C.A. caspase 3 modified mRNA fully modified with 5-methylcytidine and 1-methylpseudouridine or fully modified with 1-methylpseudouridine. FIG. 7B shows protein from C.A. caspase 6 modified mRNA fully modified with 5-methylcytidine and 1-methylpseudouridine or fully modified with 1-methylpseudouridine.

DETAILED DESCRIPTION

It is of great interest in the fields of therapeutics, diagnostics, reagents and for biological assays to be able to deliver a nucleic acid, e.g., a ribonucleic acid (RNA) inside a cell, whether in vitro, in vivo, in situ or ex vivo, such as to cause intracellular translation of the nucleic acid and production of an encoded polypeptide of interest. Of particular importance is the delivery and function of a non-integrative polynucleotide.

Described herein are compositions (including pharmaceutical compositions) and methods for the design, preparation, manufacture and/or formulation of polynucleotides encoding one or more polypeptides of interest. Also provided are systems, processes, devices and kits for the selection, design and/or utilization of the polynucleotides encoding the polypeptides of interest described herein.

To this end, polypeptides of the present invention are encoded by a new class of polynucleotide therapeutics, termed “signal-sensor polynucleotides” which are particularly useful in the stratification, profiling and/or personalization of the polynucleotide therapeutice (e.g., mRNA) and which are tailored to a particular cell type, disease or cell microenvironment or biological profile.

It is known that cancers exhibit diverse gene expression patterns, physicochemical environments and metastatic or motility behaviors and according to Hanahan and Weinberg (Cell, 2011, 144:646-674) there are six hallmarks of cancer. These include sustaining a proliferative signaling, evading growth suppressors, resisting cell death, enabling replicative immortality, inducing angiogenesis, and activating invasion and metastasis. These hallmarks or functions of cancer allow the cancer to survive, proliferate and disseminate and each arises at different times and in different patterns depending on the cancer type.

The development of cancer therapeutics which to selectively target the cancer cells while sparing normal cells dominates ongoing efforts in every area of oncology. The polynucleotides of the present invention represent such therapeutics; having the ability to selectively stabilize or destabilize cell systems, signal proliferation (survival) or death, trigger the cell cycle or senescence and/or activate or avoid the immune response depending on the cell type, e.g., cancer or normal cell.

According to the present invention, signal-sensor polynucleotide therapeutics may be used to destabilize the survival advantages or hallmarks of a cancer cell (hence they would be cytotoxic). In one embodiment diagnostic efforts would include the profiling of the cancer (although this would not be required a priori) including metabolic state (hypoxic, acidotic), apoptotic vs. survival gene profiles, cell cycle vs. senescent stage, immune status, and stromal factors present.

In one embodiment the signal-sensor polynucleotide disrupts the transcriptome of the cancer cell. The disruption may affect one or more signaling or expression events. For example the encoded oncology-related polypeptide may act upstream of a transcription factor known to induce or enhance the expression of genes associated with a cancer. Delivery of the signal-sensor polynucleotide encoding the oncology-related polypeptide which inhibits such a transcription factor (either by binding or sequestration or degradation) would thereby alter the transcriptome of the cancer cell and have a therapeutic benefit. One such transcription factor is HIF-1alpha. A signal-sensor polynucleotide encoding a protein which is capable of binding HIF-1alpha or whose expression results in lower HIF-1alpha, would effectively turn down HIF-1alpha regulated genes, e.g., VEGFA or SLC2A1, and destabilize the cancer.

In one embodiment, the profile of the cancer may be evaluated before the signal-sensor polynucleotide is selected. Such profiling data would inform the selection of which oncology-related polypeptide to be delivered. The profile of gene expression, categorized by hallmark class such as apoptosis, replicative capacity or metabolic signature would allow dynamic instability scoring for a polypeptide and an optimization of therapeutic window for the signal-sensor polynucleotide. As used herein, a “dynamic instability index” refers to a dose of signal-sensor polynucleotide sufficient to induce 50% increase of the oncology-related target protein in vitro in a cancer cell as compared to a normal matched cell.

Profiling may also be done within hallmark classes such as the distinction between caspase-dependent and caspase independent gene expression for the apoptosis class. Alternatively, profiling could be conducted across classes such as gene profiling of apoptosis, senescence (replicative capacity), and metabolic classes.

In one embodiment, the signal-sensor polynucleotides described herein may be used to reduce the expression and/or amount of a polypeptide in a cell. As a non-limiting example, MYC inhibitor A, MYC inhibitor B, MYC inhibitor C or MYC inhibitor D may be used on Hep3B cells in order to determine the potency of MYC inhibitor A, MYC inhibitor B, MYC inhibitor C or MYC inhibitor D at various concentrations (see e.g., Example 55).

In one embodiment, the signal-sensor polynucleotides described herein may direct either cytotoxic or cytoprotective therapeutic benefit to specific cells, e.g., normal vs. cancerous.

In one embodiment signal-sensor polynucleotides would not only encode an oncology-related polypeptide but also a sensor sequence. Sensor sequences include, for example, microRNA binding sites, transcription factor binding sites, artificial binding sites engineered to act as pseudo-receptors for endogenous nucleic acid binding molecules. A “sensor region” is a region of linked nucleosides of the signal-sensor polynucleotide comprising at least one sensor sequence. The signal-sensor polynucleotides of the present invention may have one or more sensor regions.

In one embodiment, one or more sensor regions may be located in the first flanking region. As a non-limiting example, the sensor region in the first flanking region may comprise at least one sensor sequence. The sensor sequence may be, but is not limited to, mir-122, mir-142-3p, mir-142-5p, mir-146, fragments or variants thereof. As another non-limiting example, the sensor region in the first flanking region may comprise at least one sensor sequence such as a mir-122 sequence. The mir-122 sequence may be, but is not limited to, a mir-122 binding site, mir-122 seed sequence, mir-122 binding site without the seed sequence or a combination thereof.

In another embodiment, one or more sensor regions may be located in the second flanking region. As a non-limiting example, the sensor region in the second flanking region may include a sensor sequence such as mir-122, mir-142-3p, mir-142-5p, mir-146, fragments or variants thereof. As another non-limiting example, the sensor region in the second flanking region may include three sensor sequences. The sensor sequences may be, but are not limited to, mir-122 sequences such as mir-122 binding sites, mir-122 seed sequences, mir-122 binding sites without the seed sequence or a combination thereof. As yet another non-limiting example, the sensor region in the second flanking region is located in the 3′UTR and the sensor region may include a sensor sequence which is a mir-122 sequence. The mir-122 sequence may be, but is not limited to, a mir-122 binding site, mir-122 seed sequence, mir-122 binding site without the seed sequence or a combination thereof.

In one embodiment, two or more sensor regions may be located in the same region of the signal-sensor polynucleotide such as, but not limited to, a first region first region of linked nucleotides, the first flanking region and/or the second flanking region. As a non-limiting example, the two or more sensor regions are located in the second flanking region. As yet another non-limiting example, three sensor regions are located in the 3′ UTR in the second flanking region. The three sensor regions may include, mir-122 binding sites, mir-122 seed sequences, mir-122 binding sites without the seed sequence or a combination thereof.

In another embodiment, two or more sensor regions may be located in different regions of the signal-sensor polynucleotide such as, but not limited to, the first region of linked nucleotides, the first flanking region and/or the second flanking region. As a non-limiting example, a first sensor region is located in the first flanking region and a second sensor region is located in the second flanking region. The sensor regions may comprise the same sensor sequence or different sensor sequences.

In one embodiment, a start codon is located within a sensor region.

In one embodiment, a sensor region may comprise two or more sensor sequences. The sensor sequences may be the same or different.

In one embodiment, the sensor region may comprise two or more sensor sequence which are different from each other but they may be based on the same mir binding site. As a non-limiting example, the sensor region may include at least one miR binding site sequence and at least one mir binding site sequence with the seed removed. As another non-limiting example, the sensor region may include at least one miR binding site sequence and at least one miR seed sequence. As yet another non-limiting example, the sensor region may include at least one miR binding site sequence with the seed removed and at least one miR seed sequence.

In another embodiment, the sensor region may comprise two or more sensor sequences which are in a pattern such as ABABAB or AABBAABBAABB or ABCABCABC or variants thereof repeated once, twice, or more than three times. In these patterns, each letter, A, B, or C represent a different miR sequence.

In yet another embodiment, the signal-sensor polynucleotide may include two or more sensor regions with each sensor region having one or more sensor sequences. As a non-limiting example, the sensor sequences may be in a pattern such as ABABAB or AABBAABBAABB or ABCABCABC or variants thereof repeated once, twice, or more than three times in each of the sensor regions. As another non-limiting example, the sensor sequences may be in a pattern such as ABABAB or AABBAABBAABB or ABCABCABC or variants thereof repeated once, twice, or more than three times across the entire signal-sensor polynucleotide. In these patterns, each letter, A, B, or C represent a different miR sequence. As a non-limiting example, the first sensor region may have sensor sequences in the pattern ABA and the second sensor region may have sensor sequences in the pattern BAB so the overall pattern of the sensor sequences in the signal-sensor polynucleotide is ABABAB. As another non-limiting example, the first sensor region may have sensor sequences AA, the second sensor region may have sensor sequences BB, the third sensor region may have sensor sequences AA and the fourth sensor region may have sensor sequences BB so the overall pattern of the sensor sequences in the signal-sensor polynucleotide is AABBAABB.

The sensor sequences in the signal-sensor polynucleotides of the present invention may include one or more regulatory sequences in the 3-UTR and/or 5′UTR of natural mRNAs, which regulate mRNA stability and translation in different tissues and cells. Such cis-regulatory elements may include, but are not limited to, Cis-RNP (Ribonucleoprotein)/RBP (RNA binding protein) regulatory elements, AU-rich element AUE, structured stem-loop, constitutive decay elements (CDEs), GC-richness and other structured mRNA motifs (Parker B J et al., Genome Research, 2011, 21, 1929-1943, which is herein incorporated by reference in its entirety.). For example, CDEs are a class of regulatory motifs that mediate mRNA degradation through their interaction with Roquin proteins. In particular, CDEs are found in many mRNAs that encode regulators of development and inflammation to limit cytokine production in macrophage (Leppek K et al., Cell, 2013, 153, 869-881, which is herein incorporated by reference in its entirety.).

In one embodiment, a particular CDE can be introduced to the signal-sensor polynucleotide when the degradation of polypeptides in a cell or tissue is desired. A particular CDE can also be removed from the signal-sensor polynucleotide in order to maintain a more stable mRNA in a cell or tissue for sustaining protein expression.

In one embodiment, microRNA (miRNA) profiling of the cancer cells or tissues may be conducted to determine the presence or absence of miRNA in the cells or tissues to determine the appropriate microRNA to use as sensor sequences in the signal sensor polynucleotides.

MicroRNA gene regulation may be influenced by the sequence surrounding the microRNA such as, but not limited to, the species of the surrounding sequence, the type of sequence (e.g., heterologous, homologous and artificial), regulatory elements in the surrounding sequence and/or structural elements in the surrounding sequence. The microRNA may be influenced by the 5′UTR and/or the 3′UTR. As a non-limiting example, a non-human 3′UTR may increase the regulatory effect of the microRNA sequence on the expression of a polypeptide of interest compared to a human 3′UTR of the same sequence type.

Other regulatory elements and/or structural elements of the 5′-UTR can influence microRNA mediated gene regulation. One such example is a structured IRES (Internal Ribosome Entry Site) in the 5′UTR, which is necessary for the binding of translational elongation factors to initiate protein translation. EIF4A2 binding to this secondarily structured element in the 5′UTR is necessary for microRNA mediated gene expression (Meijer H A et al., Science, 2013, 340, 82-85, herein incorporated by reference in its entirety). The sensor-signal polynucleotide can further be modified to include this structured 5′-UTR in order to enhance microRNA mediated gene regulation.

At least one microRNA site can be engineered into the 3′ UTR of the signal-sensor polynucleotides of the present invention. In this context, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten or more microRNA sites may be engineered into the 3′ UTR of the signal-sensor polynucleotides of the present invention. In one embodiment, the microRNA sites incorporated into the signal-sensor polynucleotides may be the same or may be different microRNA sites. In another embodiment, the microRNA sites incorporated into the signal-sensor polynucleotides may target the same or different tissues in the body. As a non-limiting example, through the introduction of tissue-, cell-type-, or disease-specific microRNA binding sites in the 3′ UTR of a signal-sensor polynucleotide, the degree of expression in specific cell types (e.g. hepatocytes, myeloid cells, endothelial cells, cancer cells, etc.) can be reduced.

In one embodiment, a microRNA site can be engineered near the 5′ terminus of the 3′UTR, about halfway between the 5′ terminus and 3′terminus of the 3′UTR and/or near the 3′terminus of the 3′UTR. As a non-limiting example, a microRNA site may be engineered near the 5′ terminus of the 3′UTR and about halfway between the 5′ terminus and 3′terminus of the 3′UTR. As another non-limiting example, a microRNA site may be engineered near the 3′terminus of the 3′UTR and about halfway between the 5′ terminus and 3′terminus of the 3′UTR. As yet another non-limiting example, a microRNA site may be engineered near the 5′ terminus of the 3′UTR and near the 3′ terminus of the 3′UTR.

In another embodiment, a 3′UTR can comprise 4 microRNA sites. The microRNA sites may be complete microRNA binding sites, microRNA seed sequences and/or microRNA binding site sequences without the seed sequence.

In one embodiment, a signal-sensor polynucleotide may be engineered to include microRNA sites which are expressed in different tissues of a subject. As a non-limiting example, a signal-sensor polynucleotide of the present invention may be engineered to include miR-192 and miR-122 to regulate expression of the signal-sensor polynucleotide in the liver and kidneys of a subject. In another embodiment, a signal-sensor polynucleotide may be engineered to include more than one microRNA sites for the same tissue. For example a signal-sensor polynucleotide of the present invention may be engineered to include miR-17-92 and miR-126 to regulate expression of the signal-sensor polynucleotide in endothelial cells of a subject.

In one embodiment, the therapeutic window and or differential expression associated with the oncology-related polypeptide encoded by the signal-sensor polynucleotide of the invention may be altered. For example, signal-sensor polynucleotides may be designed whereby a death signal is more highly expressed in cancer cells (or a survival signal in a normal cell) by virtue of the miRNA signature of those cells. Where a cancer cell expresses a lower level of a particular miRNA, the signal-sensor polynucleotide encoding the binding site for that miRNA (or miRNAs) would be more highly expressed. Hence, the oncology-related polypeptide encoded by the signal-sensor polynucleotide is selected as a protein which triggers or induces cell death. Neighboring noncancer cells, harboring a higher expression of the same miRNA would be less affected by the encoded death signal as the signal-sensor polynucleotide would be expressed at a lower level due to the affects of the miRNA binding to the binding site or “sensor” encoded in the 3′UTR. Conversely, cell survival or cytoprotective signals may be delivered to tissues containing cancer and non cancerous cells where a miRNA has a higher expression in the cancer cells—the result being a lower survival signal to the cancer cell and a larger survival signature to the normal cell. Multiple signal-sensor polynucleotides may be designed and administered having different signals according to the previous paradigm.

In one embodiment, the expression of a signal-sensor polynucleotide may be controlled by incorporating at least one sensor sequence in the signal-sensor polynucleotide and formulating the signal-sensor polynucleotide. As a non-limiting example, a polynucleotide may be targeted to an orthotopic tumor by having a polynucleotide incorporating a miR-122 binding site and formulated in a lipid nanoparticle comprising the cationic lipid DLin-KC2-DMA (see e.g., the experiments described in Example 56A and 56B).

Through an understanding of the expression patterns of microRNA in different cell types, signal-sensor polynucleotides can be engineered for more targeted expression in specific cell types or only under specific biological conditions. Through introduction of tissue-specific microRNA binding sites, signal-sensor polynucleotides could be designed that would be optimal for protein expression in a tissue or in the context of a biological condition such as cancer.

Transfection experiments can be conducted in relevant cell lines, using engineered signal-sensor polynucleotides and protein production can be assayed at various time points post-transfection. For example, cells can be transfected with different microRNA binding site-engineering nucleic acids or signal-sensor polynucleotides and by using an ELISA kit to the relevant protein and assaying protein produced at 6 hr, 12 hr, 24 hr, 48 hr, 72 hr and 7 days post-transfection. In vivo experiments can also be conducted using microRNA-binding site-engineered molecules to examine changes in tissue-specific expression of formulated signal-sensor polynucleotides.

In one embodiment, the signal-sensor polynucleotides of the invention may include at least one microRNA in order to dampen the antigen presentation by antigen presenting cells. The microRNA may be the complete microRNA sequence, the microRNA seed sequence, the microRNA sequence without the seed or a combination thereof. As a non-limiting example, the microRNA incorporated into the signal-sensor polynucleotide may be specific to the hematopoietic system. As another non-limiting example, the microRNA incorporated into the signal-sensor polynucleotides of the invention to dampen antigen presentation is miR-142-3p.

In one embodiment, the signal-sensor polynucleotides of the invention may include at least one microRNA in order to dampen expression of the encoded polypeptide in a cell of interest. As a non-limiting example, the signal-sensor polynucleotides of the invention may include at least one miR-122 binding site in order to dampen expression of an encoded polypeptide of interest in the liver. As another non-limiting example, the signal-sensor polynucleotides of the invention may include at least one miR-142-3p binding site, miR-142-3p seed sequence, miR-142-3p binding site without the seed, miR-142-5p binding site, miR-142-5p seed sequence, miR-142-5p binding site without the seed, miR-146 binding site, miR-146 seed sequence and/or miR-146 binding site without the seed sequence (see e.g., the experiment outlined in Example 47 and Example 60).

According to the present invention, the signal-sensor polynucleotides described herein may be modified as to avoid the deficiencies of other polypeptide-encoding molecules of the art. Hence, in this embodiment the signal-sensor polynucleotides are referred to as modified signal-sensor polynucleotides or primary constructs, modified mRNA or mmRNA.

Provided herein, in part, are signal-sensor polynucleotide polynucleotides, primary constructs and/or mmRNA encoding oncology-related polypeptides of interest which have been designed to improve one or more of the stability and/or clearance in tissues, receptor uptake and/or kinetics, cellular access by the compositions, engagement with translational machinery, mRNA half-life, translation efficiency, immune evasion, protein production capacity, secretion efficiency (when applicable), accessibility to circulation, protein half-life and/or modulation of a cell's status, function and/or activity.

I. COMPOSITIONS OF THE INVENTION

The present invention provides nucleic acid molecules, specifically signal-sensor polynucleotides, primary constructs and/or mmRNA which encode one or more oncology-related polypeptides of interest. Specifically the invention contemplates signal-sensor polynucleotides which are useful in cancer or cancer related diseases, disorders. As used herein, “signal-sensor polynucleotides” are nucleic acid transcripts which encode one or more oncology-related polypeptides of interest that, when translated, delivers a “signal” to the cell (cancer or noncancerous) which results in the therapeutic benefit to the organism of either being detrimental to the cancer cell or beneficial to normal cells or both detrimental to cancer cells and advantageous to normal cells. The signal-sensor polynucleotides may optionally further comprise a sequence (translatable or not) which “senses” the microenvironment of the polynucleotide and alters (a) the function or phenotypic outcome associated with the peptide or protein which is translated, (b) the expression level of the signal-sensor polynucleotide, and/or both.

The term “nucleic acid,” in its broadest sense, includes any compound and/or substance that comprise a polymer of nucleotides. These polymers are often referred to as polynucleotides. Exemplary nucleic acids or polynucleotides of the invention include, but are not limited to, ribonucleic acids (RNAs), deoxyribonucleic acids (DNAs), threose nucleic acids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs), locked nucleic acids (LNAs, including LNA having a β-D-ribo configuration, α-LNA having an α-L-ribo configuration (a diastereomer of LNA), 2′-amino-LNA having a 2′-amino functionalization, and 2′-amino-α-LNA having a 2′-amino functionalization) or hybrids thereof.

In preferred embodiments, the signal-sensor polynucleotide or nucleic acid molecule is a messenger RNA (mRNA). As used herein, the term “messenger RNA” (mRNA) refers to any polynucleotide which encodes a polypeptide of interest and which is capable of being translated to produce the encoded polypeptide of interest in vitro, in vivo, in situ or ex vivo. Signal-sensor polynucleotides of the invention may be mRNA or any nucleic acid molecule and may or may not be chemically modified.

Traditionally, the basic components of an mRNA molecule include at least a coding region, a 5′UTR, a 3′UTR, a 5′ cap and a poly-A tail. Building on this wild type modular structure, the present invention expands the scope of functionality of traditional mRNA molecules by providing signal-sensor polynucleotides or primary RNA constructs which maintain a modular organization, but which comprise one or more structural and/or chemical modifications or alterations which impart useful properties to the polynucleotide including, in some embodiments, the lack of a substantial induction of the innate immune response of a cell into which the signal-sensor polynucleotide is introduced. As such, modified mRNA molecules of the present invention, which may be synthetic, are termed “mmRNA.” As used herein, a “structural” feature or modification is one in which two or more linked nucleotides are inserted, deleted, duplicated, inverted or randomized in a signal-sensor polynucleotide polynucleotide, primary construct or mmRNA without significant chemical modification to the nucleotides themselves. Because chemical bonds will necessarily be broken and reformed to effect a structural modification, structural modifications are of a chemical nature and hence are chemical modifications. However, structural modifications will result in a different sequence of nucleotides. For example, the polynucleotide “ATCG” may be chemically modified to “AT-5meC-G”. The same polynucleotide may be structurally modified from “ATCG” to “ATCCCG”. Here, the dinucleotide “CC” has been inserted, resulting in a structural modification to the polynucleotide.

Signal-Sensor Polynucleotide, Primary Construct or mmRNA Architecture

The signal-sensor polynucleotides of the present invention are distinguished from wild type mRNA in their functional and/or structural design features which serve to, as evidenced herein, overcome existing problems of effective polypeptide production using nucleic acid-based therapeutics.

FIG. 1 shows a representative signal-sensor primary construct 100 of the present invention. As used herein, the term “primary construct” or “primary mRNA construct” refers to a signal-sensor polynucleotide transcript which encodes one or more polypeptides of interest and which retains sufficient structural and/or chemical features to allow the polypeptide of interest encoded therein to be translated. Signal-sensor primary constructs may be polynucleotides of the invention. When structurally or chemically modified, the signal-sensor primary construct may be referred to as a mmRNA.

Returning to FIG. 1, the primary construct 100 here contains a first region of linked nucleotides 102 that is flanked by a first flanking region 104 and a second flaking region 106. As used herein, the “first region” may be referred to as a “coding region” or “region encoding” or simply the “first region.” This first region may include, but is not limited to, the encoded oncology-related polypeptide of interest. The oncology-related polypeptide of interest may comprise at its 5′ terminus one or more signal peptide sequences encoded by a signal peptide sequence region 103. The flanking region 104 may comprise a region of linked nucleotides comprising one or more complete or incomplete 5′ UTRs sequences. The flanking region 104 may also comprise a 5′ terminal cap 108. The second flanking region 106 may comprise a region of linked nucleotides comprising one or more complete or incomplete 3′ UTRs. The flanking region 106 may also comprise a 3′ tailing sequence 110 and a 3′UTR 120.

Bridging the 5′ terminus of the first region 102 and the first flanking region 104 is a first operational region 105. Traditionally this operational region comprises a start codon. The operational region may alternatively comprise any translation initiation sequence or signal including a start codon.

Bridging the 3′ terminus of the first region 102 and the second flanking region 106 is a second operational region 107. Traditionally this operational region comprises a stop codon. The operational region may alternatively comprise any translation initiation sequence or signal including a stop codon. According to the present invention, multiple serial stop codons may also be used. In one embodiment, the operation region of the present invention may comprise two stop codons. The first stop codon may be “TGA” and the second stop codon may be selected from the group consisting of “TAA,” “TGA” and “TAG.” The operation region may further comprise three stop codons. The third stop codon may be selected from the group consisting of “TAA,” “TGA” and “TAG.”

Turning to FIG. 2, the 3′UTR 120 of the second flanking region 106 may comprise one or more sensor sequences 130. A region comprising at least one sensor sequence is referred to as a “sensor region.” These sensor sequences as discussed herein operate as pseudo-receptors (or binding sites) for ligands of the local microenvironment of the primary construct or signal-sensor polynucleotide. For example, microRNA binding sites or miRNA seeds may be used as sensors such that they function as pseudoreceptors for any microRNAs present in the environment of the polynucleotide.

Generally, the shortest length of the first region of the signal-sensor primary construct of the present invention can be the length of a nucleic acid sequence that is sufficient to encode for a dipeptide, a tripeptide, a tetrapeptide, a pentapeptide, a hexapeptide, a heptapeptide, an octapeptide, a nonapeptide, or a decapeptide. In another embodiment, the length may be sufficient to encode a peptide of 2-30 amino acids, e.g. 5-30, 10-30, 2-25, 5-25, 10-25, or 10-20 amino acids. The length may be sufficient to encode for a peptide of at least 11, 12, 13, 14, 15, 17, 20, 25 or 30 amino acids, or a peptide that is no longer than 40 amino acids, e.g. no longer than 35, 30, 25, 20, 17, 15, 14, 13, 12, 11 or 10 amino acids. Examples of dipeptides that the polynucleotide sequences can encode or include, but are not limited to, carnosine and anserine.

Generally, the length of the first region encoding the oncology-related polypeptide of interest of the present invention is greater than about 30 nucleotides in length (e.g., at least or greater than about 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,500, and 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, 10,000, 20,000, 30,000, 40,000, 50,000, 60,000, 70,000, 80,000, 90,000 or up to and including 100,000 nucleotides). As used herein, the “first region” may be referred to as a “coding region” or “region encoding” or simply the “first region.”

In some embodiments, the signal-sensor polynucleotide, primary construct, or mmRNA includes from about 30 to about 100,000 nucleotides (e.g., from 30 to 50, from 30 to 100, from 30 to 250, from 30 to 500, from 30 to 1,000, from 30 to 1,500, from 30 to 3,000, from 30 to 5,000, from 30 to 7,000, from 30 to 10,000, from 30 to 25,000, from 30 to 50,000, from 30 to 70,000, from 100 to 250, from 100 to 500, from 100 to 1,000, from 100 to 1,500, from 100 to 3,000, from 100 to 5,000, from 100 to 7,000, from 100 to 10,000, from 100 to 25,000, from 100 to 50,000, from 100 to 70,000, from 100 to 100,000, from 500 to 1,000, from 500 to 1,500, from 500 to 2,000, from 500 to 3,000, from 500 to 5,000, from 500 to 7,000, from 500 to 10,000, from 500 to 25,000, from 500 to 50,000, from 500 to 70,000, from 500 to 100,000, from 1,000 to 1,500, from 1,000 to 2,000, from 1,000 to 3,000, from 1,000 to 5,000, from 1,000 to 7,000, from 1,000 to 10,000, from 1,000 to 25,000, from 1,000 to 50,000, from 1,000 to 70,000, from 1,000 to 100,000, from 1,500 to 3,000, from 1,500 to 5,000, from 1,500 to 7,000, from 1,500 to 10,000, from 1,500 to 25,000, from 1,500 to 50,000, from 1,500 to 70,000, from 1,500 to 100,000, from 2,000 to 3,000, from 2,000 to 5,000, from 2,000 to 7,000, from 2,000 to 10,000, from 2,000 to 25,000, from 2,000 to 50,000, from 2,000 to 70,000, and from 2,000 to 100,000).

According to the present invention, the first and second flanking regions may range independently from 15-1,000 nucleotides in length (e.g., greater than 30, 40, 45, 50, 55, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, and 900 nucleotides or at least 30, 40, 45, 50, 55, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, and 1,000 nucleotides).

According to the present invention, the tailing sequence may range from absent to 500 nucleotides in length (e.g., at least 60, 70, 80, 90, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, or 500 nucleotides). Where the tailing region is a polyA tail, the length may be determined in units of or as a function of polyA binding protein binding. In this embodiment, the polyA tail is long enough to bind at least 4 monomers of polyA binding protein. PolyA binding protein monomers bind to stretches of approximately 38 nucleotides. As such, it has been observed that polyA tails of about 80 nucleotides and 160 nucleotides are functional.

According to the present invention, the capping region may comprise a single cap or a series of nucleotides forming the cap. In this embodiment the capping region may be from 1 to 10, e.g. 2-9, 3-8, 4-7, 1-5, 5-10, or at least 2, or 10 or fewer nucleotides in length. In some embodiments, the cap is absent.

According to the present invention, the first and second operational regions may range from 3 to 40, e.g., 5-30, 10-20, 15, or at least 4, or 30 or fewer nucleotides in length and may comprise, in addition to a start and/or stop codon, one or more signal and/or restriction sequences.

Cyclic Signal-Sensor Polynucleotides

According to the present invention, a signal-sensor primary construct or mmRNA may be cyclized, or concatemerized, to generate a translation competent molecule to assist interactions between poly-A binding proteins and 5′-end binding proteins. The mechanism of cyclization or concatemerization may occur through at least 3 different routes: 1) chemical, 2) enzymatic, and 3) ribozyme catalyzed. The newly formed 5′-/3′-linkage may be intramolecular or intermolecular.

In the first route, the 5′-end and the 3′-end of the nucleic acid may contain chemically reactive groups that, when close together, form a new covalent linkage between the 5′-end and the 3′-end of the molecule. The 5′-end may contain an NHS-ester reactive group and the 3′-end may contain a 3′-amino-terminated nucleotide such that in an organic solvent the 3′-amino-terminated nucleotide on the 3′-end of a synthetic mRNA molecule will undergo a nucleophilic attack on the 5′-NHS-ester moiety forming a new 5′-/3′-amide bond.

In the second route, T4 RNA ligase may be used to enzymatically link a 5′-phosphorylated nucleic acid molecule to the 3′-hydroxyl group of a nucleic acid forming a new phosphorodiester linkage. In an example reaction, 1 μg of a nucleic acid molecule is incubated at 37° C. for 1 hour with 1-10 units of T4 RNA ligase (New England Biolabs, Ipswich, Mass.) according to the manufacturer's protocol. The ligation reaction may occur in the presence of a split oligonucleotide capable of base-pairing with both the 5′- and 3′-region in juxtaposition to assist the enzymatic ligation reaction.

In the third route, either the 5′- or 3′-end of the cDNA template encodes a ligase ribozyme sequence such that during in vitro transcription, the resultant nucleic acid molecule can contain an active ribozyme sequence capable of ligating the 5′-end of a nucleic acid molecule to the 3′-end of a nucleic acid molecule. The ligase ribozyme may be derived from the Group I Intron, Group I Intron, Hepatitis Delta Virus, Hairpin ribozyme or may be selected by SELEX (systematic evolution of ligands by exponential enrichment). The ribozyme ligase reaction may take 1 to 24 hours at temperatures between 0 and 37° C.

Signal-Sensor Polynucleotide Multimers

According to the present invention, multiple distinct signal-sensor polynucleotides, primary constructs or mmRNA may be linked together through the 3′-end using nucleotides which are modified at the 3′-terminus. Chemical conjugation may be used to control the stoichiometry of delivery into cells. For example, the glyoxylate cycle enzymes, isocitrate lyase and malate synthase, may be supplied into HepG2 cells at a 1:1 ratio to alter cellular fatty acid metabolism. This ratio may be controlled by chemically linking signal-sensor polynucleotides, primary constructs or mmRNA using a 3′-azido terminated nucleotide on one signal-sensor polynucleotide, primary construct or mmRNA species and a C5-ethynyl or alkynyl-containing nucleotide on the opposite signal-sensor polynucleotide, primary construct or mmRNA species. The modified nucleotide is added post-transcriptionally using terminal transferase (New England Biolabs, Ipswich, Mass.) according to the manufacturer's protocol. After the addition of the 3′-modified nucleotide, the two signal-sensor polynucleotide, primary construct or mmRNA species may be combined in an aqueous solution, in the presence or absence of copper, to form a new covalent linkage via a click chemistry mechanism as described in the literature.

In another example, more than two signal-sensor polynucleotides may be linked together using a functionalized linker molecule. For example, a functionalized saccharide molecule may be chemically modified to contain multiple chemical reactive groups (SH—, NH2—, N3, etc. . . . ) to react with the cognate moiety on a 3′-functionalized signal-sensorpolynucleotide molecule (i.e., a 3′-maleimide ester, 3′-NHS-ester, alkynyl). The number of reactive groups on the modified saccharide can be controlled in a stoichiometric fashion to directly control the stoichiometric ratio of conjugated signal-sensor polynucleotide, primary construct or mmRNA.

Signal-Sensor Polynucleotide Conjugates and Combinations

In order to further enhance oncology-related protein production, signal-sensor polynucleotide primary constructs or mmRNA of the present invention can be designed to be conjugated to other polynucleotides, oncology-related polypeptides, dyes, intercalating agents (e.g. acridines), cross-linkers (e.g. psoralene, mitomycin C), porphyrins (TPPC4, texaphyrin, Sapphyrin), polycyclic aromatic hydrocarbons (e.g., phenazine, dihydrophenazine), artificial endonucleases (e.g. EDTA), alkylating agents, phosphate, amino, mercapto, PEG (e.g., PEG-40K), MPEG, [MPEG]2, polyamino, alkyl, substituted alkyl, radiolabeled markers, enzymes, haptens (e.g. biotin), transport/absorption facilitators (e.g., aspirin, vitamin E, folic acid), synthetic ribonucleases, proteins, e.g., glycoproteins, or peptides, e.g., molecules having a specific affinity for a co-ligand, or antibodies e.g., an antibody, that binds to a specified cell type such as a cancer cell, endothelial cell, or bone cell, hormones and hormone receptors, non-peptidic species, such as lipids, lectins, carbohydrates, vitamins, cofactors, or a drug.

Conjugation may result in increased stability and/or half life and may be particularly useful in targeting the signal-sensor polynucleotides, primary constructs or mmRNA to specific sites in the cell, tissue or organism.

According to the present invention, the signal-sensor polynucleotide mmRNA or primary constructs may be administered with, or further encode one or more of RNAi agents, siRNAs, shRNAs, miRNAs, miRNA binding sites, antisense RNAs, ribozymes, catalytic DNA, tRNA, RNAs that induce triple helix formation, aptamers or vectors, and the like.

In one embodiment, the signal-sensor polynucleotides described herein may be conjugated with a moiety to target various cancer cells such as, but not limited to, the moieties described in US Patent Application No. US20130216561, the contents of which are herein incorporated by reference in its entirety. The linkage between the signal-sensor polynucleotides and the cancer targeting moiety may be an acid cleavable linkage that can increase the efficacy of the conjugate such as, but not limited to, the linkages described in US Patent Application No. US20130216561, the contents of which are herein incorporated by reference in its entirety.

Bifunctional Signal-Sensor Polynucleotide

In one embodiment of the invention are bifunctional signal-sensor polynucleotides (e.g., bifunctional primary constructs or bifunctional mmRNA). As the name implies, bifunctional signal-sensor polynucleotides are those having or capable of at least two functions. These molecules may also by convention be referred to as multi-functional.

The multiple functionalities of bifunctional signal-sensor polynucleotides may be encoded by the RNA (the function may not manifest until the encoded product is translated) or may be a property of the polynucleotide itself. It may be structural or chemical. Bifunctional modified signal-sensor polynucleotides may comprise a function that is covalently or electrostatically associated with the polynucleotides. Further, the two functions may be provided in the context of a complex of a signal-sensor polynucleotide and another molecule.

Bifunctional signal-sensor polynucleotides may encode oncology-related peptides which are anti-proliferative. These peptides may be linear, cyclic, constrained or random coil. They may function as aptamers, signaling molecules, ligands or mimics or mimetics thereof. Anti-proliferative peptides may, as translated, be from 3 to 50 amino acids in length. They may be 5-40, 10-30, or approximately 15 amino acids long. They may be single chain, multichain or branched and may form complexes, aggregates or any multi-unit structure once translated.

Noncoding Signal-Sensor Polynucleotides

As described herein, provided are signal-sensor polynucleotides and primary constructs having sequences that are partially or substantially not translatable, e.g., having a noncoding region. Such noncoding region may be the “first region” of the signal-sensor primary construct. Alternatively, the noncoding region may be a region other than the first region. Such molecules are generally not translated, but can exert an effect on protein production by one or more of binding to and sequestering one or more translational machinery components such as a ribosomal protein or a transfer RNA (tRNA), thereby effectively reducing protein expression in the cell or modulating one or more pathways or cascades in a cell which in turn alters protein levels. The signal-sensor polynucleotide and/or primary construct may contain or encode one or more long noncoding RNA (lncRNA, or lincRNA) or portion thereof, a small nucleolar RNA (sno-RNA), micro RNA (miRNA), small interfering RNA (siRNA) or Piwi-interacting RNA (piRNA).

Auxotrophic Signal-Sensor Polynucleotides

In one embodiment, the signal-sensor polynucleotides of the present invention may be auxotrophic. As used herein, the term “auxotrophic” refers to signal-sensor polynucleotides that comprise at least one feature that triggers, facilitates or induces the degradation or inactivation of the itself in response to spatial or temporal cues such that oncology-related protein expression is substantially prevented or reduced. Such spatial or temporal cues include the location of the signal-sensor polynucleotide to be translated such as a particular tissue or organ or cellular environment. Also contemplated are cues involving temperature, pH, ionic strength, moisture content, and the like.

In one embodiment, the feature is located in a terminal region of the signal-sensor polynucleotides of the present invention. As a non-limiting example, the auxotrophic mRNA may contain a miR binding site in the terminal region which binds to a miR expressed in a selected tissue so that the expression of the auxotrophic mRNA is substantially prevented or reduced in the selected tissue. To this end and for example, an auxotrophic mRNA containing a miR-122 binding site will not produce protein if localized to the liver since miR-122 is expressed in the liver and binding of the miR would effectuate destruction of the auxotrophic mRNA. As a non-limiting example, HEK293 cells do not express miR-122 so there would be little to no downregulation of a signal-sensor polynucleotide having a miR-122 sequence in HEK293 but for hepatocytes which do expression miR-122 there would be a downregulation of a signal-sensor polynucleotide having a miR-122 sequence in hepatocytes (see e.g., the study outlined Example 19). As another non-limiting example, the miR-122 level can be measured in HeLa cells, primary human hepatocytes and primary rat hepatocytes prior to administration with a signal-sensor polynucleotide encoding having at least one miR-122 binding site, miR-122 binding site without the seed sequence or a miR-122 binding site After administration the expression of the signal-sensor polynucleotide can be measured to determine the dampening effect of the miR-122 in the signal-sensor polynucleotide (see e.g., the studies outlined in Examples 41, 42, 43 57, 58 and 59). As yet another non-limiting example, the effectiveness of the miR-122 binding site, miR-122 seed or the miR-122 binding site without the seed in different 3′UTRs may be evaluated in order to determine the proper UTR for the desired outcome such as, but not limited to, the highest dampening effect (see e.g., the study outlined in Example 46).

In one embodiment, the degradation or inactivation of auxotrophic mRNA may comprise a feature responsive to a change in pH. As a non-limiting example, the auxotrophic mRNA may be triggered in an environment having a pH of between pH 4.5 to 8.0 such as at a pH of 5.0 to 6.0 or a pH of 6.0 to 6.5. The change in pH may be a change of 0.1 unit, 0.2 units, 0.3 units, 0.4 units, 0.5 units, 0.6 units, 0.7 units, 0.8 units, 0.9 units, 1.0 units, 1.1 units, 1.2 units, 1.3 units, 1.4 units, 1.5 units, 1.6 units, 1.7 units, 1.8 units, 1.9 units, 2.0 units, 2.1 units, 2.2 units, 2.3 units, 2.4 units, 2.5 units, 2.6 units, 2.7 units, 2.8 units, 2.9 units, 3.0 units, 3.1 units, 3.2 units, 3.3 units, 3.4 units, 3.5 units, 3.6 units, 3.7 units, 3.8 units, 3.9 units, 4.0 units or more.

In another embodiment, the degradation or inactivation of auxotrophic mRNA may be triggered or induced by changes in temperature. As a non-limiting example, a change of temperature from room temperature to body temperature. The change of temperature may be less than 1° C., less than 5° C., less than 10° C., less than 15° C., less than 20° C., less than 25° C. or more than 25° C.

In yet another embodiment, the degradation or inactivation of auxotrophic mRNA may be triggered or induced by a change in the levels of ions in the subject. The ions may be cations or anions such as, but not limited to, sodium ions, potassium ions, chloride ions, calcium ions, magnesium ions and/or phosphate ions.

Oncology-Related Polypeptides of Interest

According to the present invention, the signal-sensor primary construct is designed to encode one or more oncology-related polypeptides of interest or fragments thereof. An oncology-related polypeptide of interest may include, but is not limited to, whole polypeptides, a plurality of polypeptides or fragments of polypeptides, which independently may be encoded by one or more nucleic acids, a plurality of nucleic acids, fragments of nucleic acids or variants of any of the aforementioned. As used herein, the term “oncology-related polypeptides of interest” refers to any polypeptide which is selected to be encoded in the signal-sensor primary construct of the present invention. As used herein, “polypeptide” means a polymer of amino acid residues (natural or unnatural) linked together most often by peptide bonds. The term, as used herein, refers to proteins, polypeptides, and peptides of any size, structure, or function. In some instances the polypeptide encoded is smaller than about 50 amino acids and the polypeptide is then termed a peptide. If the polypeptide is a peptide, it will be at least about 2, 3, 4, or at least 5 amino acid residues long. Thus, polypeptides include gene products, naturally occurring polypeptides, synthetic polypeptides, homologs, orthologs, paralogs, fragments and other equivalents, variants, and analogs of the foregoing. A polypeptide may be a single molecule or may be a multi-molecular complex such as a dimer, trimer or tetramer. They may also comprise single chain or multichain polypeptides such as antibodies or insulin and may be associated or linked. Most commonly disulfide linkages are found in multichain polypeptides. The term polypeptide may also apply to amino acid polymers in which one or more amino acid residues are an artificial chemical analogue of a corresponding naturally occurring amino acid.

The term “polypeptide variant” refers to molecules which differ in their amino acid sequence from a native or reference sequence. The amino acid sequence variants may possess substitutions, deletions, and/or insertions at certain positions within the amino acid sequence, as compared to a native or reference sequence. Ordinarily, variants will possess at least about 50% identity (homology) to a native or reference sequence, and preferably, they will be at least about 80%, more preferably at least about 90% identical (homologous) to a native or reference sequence.

In some embodiments “variant mimics” are provided. As used herein, the term “variant mimic” is one which contains one or more amino acids which would mimic an activated sequence. For example, glutamate may serve as a mimic for phosphoro-threonine and/or phosphoro-serine. Alternatively, variant mimics may result in deactivation or in an inactivated product containing the mimic, e.g., phenylalanine may act as an inactivating substitution for tyrosine; or alanine may act as an inactivating substitution for serine.

“Homology” as it applies to amino acid sequences is defined as the percentage of residues in the candidate amino acid sequence that are identical with the residues in the amino acid sequence of a second sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent homology. Methods and computer programs for the alignment are well known in the art. It is understood that homology depends on a calculation of percent identity but may differ in value due to gaps and penalties introduced in the calculation.

By “homologs” as it applies to polypeptide sequences means the corresponding sequence of other species having substantial identity to a second sequence of a second species.

“Analogs” is meant to include polypeptide variants which differ by one or more amino acid alterations, e.g., substitutions, additions or deletions of amino acid residues that still maintain one or more of the properties of the parent or starting polypeptide.

The present invention contemplates several types of compositions which are polypeptide based including variants and derivatives. These include substitutional, insertional, deletion and covalent variants and derivatives. The term “derivative” is used synonymously with the term “variant” but generally refers to a molecule that has been modified and/or changed in any way relative to a reference molecule or starting molecule.

As such, signal-sensor polynucleotides encoding oncology-related polypeptides containing substitutions, insertions and/or additions, deletions and covalent modifications with respect to reference sequences, in particular the oncology-related polypeptide sequences disclosed herein, are included within the scope of this invention. For example, sequence tags or amino acids, such as one or more lysines, can be added to the peptide sequences of the invention (e.g., at the N-terminal or C-terminal ends). Sequence tags can be used for peptide purification or localization. Lysines can be used to increase peptide solubility or to allow for biotinylation. Alternatively, amino acid residues located at the carboxy and amino terminal regions of the amino acid sequence of a peptide or protein may optionally be deleted providing for truncated sequences. Certain amino acids (e.g., C-terminal or N-terminal residues) may alternatively be deleted depending on the use of the sequence, as for example, expression of the sequence as part of a larger sequence which is soluble, or linked to a solid support.

“Substitutional variants” when referring to polypeptides are those that have at least one amino acid residue in a native or starting sequence removed and a different amino acid inserted in its place at the same position. The substitutions may be single, where only one amino acid in the molecule has been substituted, or they may be multiple, where two or more amino acids have been substituted in the same molecule.

As used herein the term “conservative amino acid substitution” refers to the substitution of an amino acid that is normally present in the sequence with a different amino acid of similar size, charge, or polarity. Examples of conservative substitutions include the substitution of a non-polar (hydrophobic) residue such as isoleucine, valine and leucine for another non-polar residue. Likewise, examples of conservative substitutions include the substitution of one polar (hydrophilic) residue for another such as between arginine and lysine, between glutamine and asparagine, and between glycine and serine. Additionally, the substitution of a basic residue such as lysine, arginine or histidine for another, or the substitution of one acidic residue such as aspartic acid or glutamic acid for another acidic residue are additional examples of conservative substitutions. Examples of non-conservative substitutions include the substitution of a non-polar (hydrophobic) amino acid residue such as isoleucine, valine, leucine, alanine, methionine for a polar (hydrophilic) residue such as cysteine, glutamine, glutamic acid or lysine and/or a polar residue for a non-polar residue.

“Insertional variants” when referring to polypeptides are those with one or more amino acids inserted immediately adjacent to an amino acid at a particular position in a native or starting sequence. “Immediately adjacent” to an amino acid means connected to either the alpha-carboxy or alpha-amino functional group of the amino acid.

“Deletional variants” when referring to polypeptides are those with one or more amino acids in the native or starting amino acid sequence removed. Ordinarily, deletional variants will have one or more amino acids deleted in a particular region of the molecule.

“Covalent derivatives” when referring to polypeptides include modifications of a native or starting protein with an organic proteinaceous or non-proteinaceous derivatizing agent, and/or post-translational modifications. Covalent modifications are traditionally introduced by reacting targeted amino acid residues of the protein with an organic derivatizing agent that is capable of reacting with selected side-chains or terminal residues, or by harnessing mechanisms of post-translational modifications that function in selected recombinant host cells. The resultant covalent derivatives are useful in programs directed at identifying residues important for biological activity, for immunoassays, or for the preparation of anti-protein antibodies for immunoaffinity purification of the recombinant glycoprotein. Such modifications are within the ordinary skill in the art and are performed without undue experimentation.

Certain post-translational modifications are the result of the action of recombinant host cells on the expressed oncology-related polypeptide. Glutaminyl and asparaginyl residues are frequently post-translationally deamidated to the corresponding glutamyl and aspartyl residues. Alternatively, these residues are deamidated under mildly acidic conditions. Either form of these residues may be present in the oncology-related polypeptides produced in accordance with the present invention.

Other post-translational modifications include hydroxylation of proline and lysine, phosphorylation of hydroxyl groups of seryl or threonyl residues, methylation of the alpha-amino groups of lysine, arginine, and histidine side chains (T. E. Creighton, Proteins: Structure and Molecular Properties, W.H. Freeman & Co., San Francisco, pp. 79-86 (1983)).

“Features” when referring to polypeptides are defined as distinct amino acid sequence-based components of a molecule. Features of the polypeptides encoded by the mmRNA of the present invention include surface manifestations, local conformational shape, folds, loops, half-loops, domains, half-domains, sites, termini or any combination thereof.

As used herein when referring to polypeptides the term “surface manifestation” refers to a polypeptide based component of a protein appearing on an outermost surface.

As used herein when referring to polypeptides the term “local conformational shape” means a polypeptide based structural manifestation of a protein which is located within a definable space of the protein.

As used herein when referring to polypeptides the term “fold” refers to the resultant conformation of an amino acid sequence upon energy minimization. A fold may occur at the secondary or tertiary level of the folding process. Examples of secondary level folds include beta sheets and alpha helices. Examples of tertiary folds include domains and regions formed due to aggregation or separation of energetic forces. Regions formed in this way include hydrophobic and hydrophilic pockets, and the like.

As used herein the term “turn” as it relates to protein conformation means a bend which alters the direction of the backbone of a peptide or polypeptide and may involve one, two, three or more amino acid residues.

As used herein when referring to polypeptides the term “loop” refers to a structural feature of a polypeptide which may serve to reverse the direction of the backbone of a peptide or polypeptide. Where the loop is found in a polypeptide and only alters the direction of the backbone, it may comprise four or more amino acid residues. Oliva et al. have identified at least 5 classes of protein loops (J. Mol Biol 266 (4): 814-830; 1997). Loops may be open or closed. Closed loops or “cyclic” loops may comprise 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids between the bridging moieties. Such bridging moieties may comprise a cysteine-cysteine bridge (Cys-Cys) typical in polypeptides having disulfide bridges or alternatively bridging moieties may be non-protein based such as the dibromozylyl agents used herein.

As used herein when referring to polypeptides the term “half-loop” refers to a portion of an identified loop having at least half the number of amino acid resides as the loop from which it is derived. It is understood that loops may not always contain an even number of amino acid residues. Therefore, in those cases where a loop contains or is identified to comprise an odd number of amino acids, a half-loop of the odd-numbered loop will comprise the whole number portion or next whole number portion of the loop (number of amino acids of the loop/2+/−0.5 amino acids). For example, a loop identified as a 7 amino acid loop could produce half-loops of 3 amino acids or 4 amino acids (7/2=3.5+/−0.5 being 3 or 4).

As used herein when referring to polypeptides the term “domain” refers to a motif of a polypeptide having one or more identifiable structural or functional characteristics or properties (e.g., binding capacity, serving as a site for protein-protein interactions).

As used herein when referring to polypeptides the term “half-domain” means a portion of an identified domain having at least half the number of amino acid resides as the domain from which it is derived. It is understood that domains may not always contain an even number of amino acid residues. Therefore, in those cases where a domain contains or is identified to comprise an odd number of amino acids, a half-domain of the odd-numbered domain will comprise the whole number portion or next whole number portion of the domain (number of amino acids of the domain/2+/−0.5 amino acids). For example, a domain identified as a 7 amino acid domain could produce half-domains of 3 amino acids or 4 amino acids (7/2=3.5+/−0.5 being 3 or 4). It is also understood that sub-domains may be identified within domains or half-domains, these subdomains possessing less than all of the structural or functional properties identified in the domains or half domains from which they were derived. It is also understood that the amino acids that comprise any of the domain types herein need not be contiguous along the backbone of the polypeptide (i.e., nonadjacent amino acids may fold structurally to produce a domain, half-domain or subdomain).

As used herein when referring to polypeptides the terms “site” as it pertains to amino acid based embodiments is used synonymously with “amino acid residue” and “amino acid side chain.” A site represents a position within a peptide or polypeptide that may be modified, manipulated, altered, derivatized or varied within the polypeptide based molecules of the present invention.

As used herein the terms “termini” or “terminus” when referring to polypeptides refers to an extremity of a peptide or polypeptide. Such extremity is not limited only to the first or final site of the peptide or polypeptide but may include additional amino acids in the terminal regions. The polypeptide based molecules of the present invention may be characterized as having both an N-terminus (terminated by an amino acid with a free amino group (NH2)) and a C-terminus (terminated by an amino acid with a free carboxyl group (COOH)). Proteins of the invention are in some cases made up of multiple polypeptide chains brought together by disulfide bonds or by non-covalent forces (multimers, oligomers). These sorts of proteins will have multiple N- and C-termini. Alternatively, the termini of the polypeptides may be modified such that they begin or end, as the case may be, with a non-polypeptide based moiety such as an organic conjugate.

Once any of the features have been identified or defined as a desired component of a polypeptide to be encoded by the signal-sensor primary construct or mmRNA of the invention, any of several manipulations and/or modifications of these features may be performed by moving, swapping, inverting, deleting, randomizing or duplicating. Furthermore, it is understood that manipulation of features may result in the same outcome as a modification to the molecules of the invention. For example, a manipulation which involved deleting a domain would result in the alteration of the length of a molecule just as modification of a nucleic acid to encode less than a full length molecule would.

Modifications and manipulations can be accomplished by methods known in the art such as, but not limited to, site directed mutagenesis. The resulting modified molecules may then be tested for activity using in vitro or in vivo assays such as those described herein or any other suitable screening assay known in the art.

According to the present invention, the oncology-related polypeptides may comprise a consensus sequence which is discovered through rounds of experimentation. As used herein a “consensus” sequence is a single sequence which represents a collective population of sequences allowing for variability at one or more sites.

As recognized by those skilled in the art, protein fragments, functional protein domains, and homologous proteins are also considered to be within the scope of oncology-related polypeptides of interest of this invention. For example, provided herein is any protein fragment (meaning an oncology-related polypeptide sequence at least one amino acid residue shorter than a reference oncology-related polypeptide sequence but otherwise identical) of a reference oncology-related protein 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or greater than 100 amino acids in length. In another example, any oncology-related protein that includes a stretch of about 20, about 30, about 40, about 50, or about 100 amino acids which are about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 100% identical to any of the sequences described herein can be utilized in accordance with the invention. In certain embodiments, a polypeptide to be utilized in accordance with the invention includes 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations as shown in any of the sequences provided or referenced herein.

Encoded Oncology-Related Polypeptides

The signal-sensor primary constructs or mmRNA of the present invention may be designed to encode oncology-related polypeptides of interest such as oncology-related peptides and proteins.

In one embodiment, signal-sensor primary constructs or mmRNA of the present invention may encode variant polypeptides which have a certain identity with a reference oncology-related polypeptide sequence. As used herein, a “reference oncology-related polypeptide sequence” refers to a starting oncology-related polypeptide sequence. Reference sequences may be wild type sequences or any sequence to which reference is made in the design of another sequence. A “reference polypeptide sequence” may, e.g., be any one of the protein sequence listed in Table 6.

The term “identity” as known in the art, refers to a relationship between the sequences of two or more peptides, as determined by comparing the sequences. In the art, identity also means the degree of sequence relatedness between peptides, as determined by the number of matches between strings of two or more amino acid residues. Identity measures the percent of identical matches between the smaller of two or more sequences with gap alignments (if any) addressed by a particular mathematical model or computer program (i.e., “algorithms”). Identity of related peptides can be readily calculated by known methods. Such methods include, but are not limited to, those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M. Stockton Press, New York, 1991; and Carillo et al., SIAM J. Applied Math. 48, 1073 (1988).

In some embodiments, the polypeptide variant may have the same or a similar activity as the reference oncology-related polypeptide. Alternatively, the variant may have an altered activity (e.g., increased or decreased) relative to a reference oncology-related polypeptide. Generally, variants of a particular signal-sensor polynucleotide or oncology-related polypeptide of the invention will have at least about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% but less than 100% sequence identity to that particular reference signal-sensor polynucleotide or oncology-related polypeptide as determined by sequence alignment programs and parameters described herein and known to those skilled in the art. Such tools for alignment include those of the BLAST suite (Stephen F. Altschul, Thomas L. Madden, Alejandro A. Schäffer, Jinghui Zhang, Zheng Zhang, Webb Miller, and David J. Lipman (1997), “Gapped BLAST and PSI-BLAST: a new generation of protein database search programs”, Nucleic Acids Res. 25:3389-3402.) Other tools are described herein, specifically in the definition of “identity.”

Default parameters in the BLAST algorithm include, for example, an expect threshold of 10, Word size of 28, Match/Mismatch Scores 1, −2, Gap costs Linear. Any filter can be applied as well as a selection for species specific repeats, e.g., Homo sapiens.

In one embodiment, the signal-sensor polynucleotides, primary constructs and/or mmRNA may be used to treat a disease, disorder and/or condition in a subject.

In one embodiment, the polynucleotides, primary constructs and/or mmRNA may be used to reduce, eliminate or prevent tumor growth in a subject.

In one embodiment, the signal-sensor polynucleotides, primary constructs and/or mmRNA may be used to reduce and/or ameliorate at least one symptom of cancer in a subject. A symptom of cancer may include, but is not limited to, weakness, aches and pains, fever, fatigue, weight loss, blood clots, increased blood calcium levels, low white blood cell count, short of breath, dizziness, headaches, hyperpigmentation, jaundice, erthema, pruritis, excessive hair growth, change in bowel habits, change in bladder function, long-lasting sores, white patches inside the mouth, white spots on the tongue, unusual bleeding or discharge, thickening or lump on parts of the body, indigestion, trouble swallowing, changes in warts or moles, change in new skin and nagging cough or hoarseness. Further, the signal-sensor polynucleotides, primary constructs and/or mmRNA may reduce a side-effect associated with cancer such as, but not limited to, chemo brain, peripheral neuropathy, fatigue, depression, nausea, vomiting, pain, anemia, lymphedema, infections, sexual side effects, reduced fertility or infertility, ostomics, insomnia and hair loss.

Oncology-Related Proteins or Oncology-Related Peptides

The signal-sensor primary constructs or mmRNA disclosed herein, may encode one or more validated or “in testing” oncology-related proteins or oncology-related peptides.

According to the present invention, one or more oncology-related proteins or oncology-related peptides currently being marketed or in development may be encoded by the oncology-related signal-sensor polynucleotide, primary constructs or mmRNA of the present invention. While not wishing to be bound by theory, it is believed that incorporation into the signal-sensor primary constructs or mmRNA of the invention will result in improved therapeutic efficacy due at least in part to the specificity, purity and selectivity of the construct designs.

The signal-sensor polynucleotides, primary constructs and/or mmRNA may alter a biological and/or physiological process and/or compound such as, but not limited to, the cell cycle, the DNA damage response (e.g., DNA damage repair), apoptosis, angiogenesis, cell motility, the epithelial to mesenchymal transition in epithelial cells, the phosphatidyl inositol 3 (PI3) kinase/Akt cellular signaling pathway, telomerase activity and/or expression, tumor metastasis, tumorigenesis, cathepsins, cell senescence, receptor tyrosine kinase signaling, metabolism and drug metabolism, G protein signaling, growth factors and receptors, heat shock proteins, histone deacetylases, hormone receptors, hypoxia, poly ADP-ribose polymerases, protein kinases, RAS signaling, topisomerases, transcription factors and tumor suppressor activity in cancerous, precancerous and/or other cells.

In one embodiment, the signal-sensor polynucleotides, primary constructs and/or mmRNA may used to express a polypeptide in cells or tissues for the purpose of replacing the protein produced from a deleted or mutated gene.

Further, the polynucleotides, primary constructs or mmRNA of the invention may be used to treat cancer which has been caused by carcinogens of natural and/or synthetic origin. In another embodiment, the use of the polynucleotides, primary constructs and/or mmRNA may be used to treat cancer caused by other organisms and/or cancers caused by viral infection.

Sensors in the Flanking Regions: Untranslated Regions (UTRs)

Untranslated regions (UTRs) of a gene are transcribed but not translated. The 5′UTR starts at the transcription start site and continues to the start codon but does not include the start codon; whereas, the 3′UTR starts immediately following the stop codon and continues until the transcriptional termination signal. There is growing body of evidence about the regulatory roles played by the UTRs in terms of stability of the nucleic acid molecule and translation. The regulatory features of a UTR can be incorporated into the signal-sensor polynucleotides, primary constructs and/or mmRNA of the present invention to enhance the stability of the molecule. The specific features can also be incorporated to ensure controlled down-regulation of the transcript in case they are misdirected to undesired organs sites. The untranslated regions may be incorporated into a vector system which can produce mRNA and/or be delivered to a cell, tissue and/or organism to produce a polypeptide of interest.

In one embodiment, the signal-sensor polynucleotides, primary constructs and/or mmRNA of the present may comprise at least one terminal modification. Non-limiting examples of terminal modifications are described in U.S. Provisional Patent Application No. 61/729,933, filed Nov. 26, 2012, entitled Terminally Optimized Modified RNAs, U.S. Provisional Patent application No. 61/737,224, filed Dec. 14, 2012, entitled Terminally Optimized RNAs, U.S. Provisional Patent Application No. 61/758,921, filed Jan. 31, 2013, entitled Differential Targeting Using RNA Constructs, U.S. Provisional Patent Application No. 61/781,139, filed Mar. 14, 2013, entitled Differential Targeting Using RNA Constructs, U.S. Provisional Patent Application No. 61/829,359, filed May 31, 2013, entitled Differential Targeting Using RNA Constructs, U.S. Provisional Patent Application No. 61/839,903, filed Jun. 27, 2013, entitled Differential Targeting Using RNA Constructs, U.S. Provisional Patent Application No. 61/842,709, filed Jul. 3, 2013, entitled Differential Targeting Using RNA Constructs, and U.S. Provisional Patent Application No. 61/857,436, filed Jul. 23, 2013, entitled Differential Targeting Using RNA Constructs, the contents of each of which are herein incorporated by reference in their entireties. These terminal modifications include, but are not limited to, 5′caps, microRNA binding sites in the terminal region, chain terminating nucleosides, translation enhancer elements in the terminal region and tailing sequences including a polyA-G quartet and stem loop sequences.

5′ UTR and Translation Initiation

Natural 5′UTRs bear features which play roles in for translation initiation. They harbor signatures like Kozak sequences which are commonly known to be involved in the process by which the ribosome initiates translation of many genes. Kozak sequences have the consensus CCR(A/G)CCAUGG, where R is a purine (adenine or guanine) three bases upstream of the start codon (AUG), which is followed by another ‘G’. 5′UTR also have been known to form secondary structures which are involved in elongation factor binding. For example, one of the secondary 5′-UTR structures is the structured IRES for eIF4A2 elongation factor binding, which is necessary for the microRNA mediated gene repression at 3′-UTR.

5′UTR secondary structures involved in elongation factor binding can interact with other RNA binding molecules in the 5′UTR or 3′UTR to regulate gene expression. For example, the elongation factor EIF4A2 binding to a secondarily structured element in the 5′UTR is necessary for microRNA mediated repression (Meijer H A et al., Science, 2013, 340, 82-85, herein incorporated by reference in its entirety). The different secondary structures in the 5′UTR can be incorporated into the flanking region to either stabilize or selectively destalized mRNAs in specific tissues or cells.

By engineering the features typically found in abundantly expressed genes of specific target organs, one can enhance the stability and oncology-related protein production of the signal-sensor polynucleotides, primary constructs or mmRNA of the invention. For example, introduction of 5′ UTR of liver-expressed mRNA, such as albumin, serum amyloid A, Apolipoprotein A/B/E, transferrin, alpha fetoprotein, erythropoietin, or Factor VIII, could be used to enhance expression of a nucleic acid molecule, such as a mmRNA, in hepatic cell lines or liver. Likewise, use of 5′ UTR from other tissue-specific mRNA to improve expression in that tissue is possible—for muscle (MyoD, Myosin, Myoglobin, Myogenin, Herculin), for endothelial cells (Tie-1, CD36), for myeloid cells (C/EBP, AML1, G-CSF, GM-CSF, CD11b, MSR, Fr-1, i-NOS), for leukocytes (CD45, CD18), for adipose tissue (CD36, GLUT4, ACRP30, adiponectin) and for lung epithelial cells (SP-A/B/C/D).

Other non-UTR sequences may be incorporated into the 5′ (or 3′ UTR) UTRs. For example, introns or portions of introns sequences may be incorporated into the flanking regions of the signal-sensor polynucleotides, primary constructs or mmRNA of the invention. Incorporation of intronic sequences may increase protein production as well as mRNA levels.

Translation Enhancer Elements (TEEs)

In one embodiment, the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA may include at least one translational enhancer polynucleotide, translation enhancer element, translational enhancer elements (collectively referred to as “TEE”s). As a non-limiting example, the TEE may be located between the transcription promoter and the start codon. The signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA with at least one TEE in the 5′UTR may include a cap at the 5′UTR. Further, at least one TEE may be located in the 5′UTR of signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA undergoing cap-dependent or cap-independent translation.

The term “translational enhancer element” or “translation enhancer element” (herein collectively referred to as “TEE”) refers to sequences that increase the amount of polypeptide or protein produced from an mRNA.

In one embodiment, TEEs are conserved elements in the UTR which can promote translational activity of a nucleic acid such as, but not limited to, cap-dependent or cap-independent translation. The conservation of these sequences has been previously shown by Panek et al (Nucleic Acids Research, 2013, 1-10; herein incorporated by reference in its entirety) across 14 species including humans.

In one embodiment, the TEE may be any of the TEEs listed in Table 35 in Example 45, including portion and/or fragments thereof. The TEE sequence may include at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99% or more than 99% of the TEE sequences disclosed in Table 35 and/or the TEE sequence may include a 5-30 nucleotide fragment, a 5-25 nucleotide fragment, a 5-20 nucleotide fragment, a 5-15 nucleotide fragment, a 5-10 nucleotide fragment of the TEE sequences disclosed in Table 35.

In one non-limiting example, the TEEs known may be in the 5′-leader of the Gtx homeodomain protein (Chappell et al., Proc. Natl. Acad. Sci. USA 101:9590-9594, 2004, herein incorporated by reference in their entirety).

In another non-limiting example, TEEs are disclosed as SEQ ID NOs: 1-35 in US Patent Publication No. US20090226470, SEQ ID NOs: 1-35 in US Patent Publication US20130177581, SEQ ID NOs: 1-35 in International Patent Publication No. WO2009075886, SEQ ID NOs: 1-5, and 7-645 in International Patent Publication No. WO2012009644, SEQ ID NO: 1 in International Patent Publication No. WO1999024595, SEQ ID NO: 1 in U.S. Pat. No. 6,310,197, and SEQ ID NO: 1 in U.S. Pat. No. 6,849,405, each of which is herein incorporated by reference in its entirety.

In yet another non-limiting example, the TEE may be an internal ribosome entry site (IRES), HCV-IRES or an IRES element such as, but not limited to, those described in U.S. Pat. No. 7,468,275, US Patent Publication Nos. US20070048776 and US20110124100 and International Patent Publication Nos. WO2007025008 and WO2001055369, each of which is herein incorporated by reference in its entirety. The IRES elements may include, but are not limited to, the Gtx sequences (e.g., Gtx9-nt, Gtx8-nt, Gtx7-nt) described by Chappell et al. (Proc. Natl. Acad. Sci. USA 101:9590-9594, 2004) and Zhou et al. (PNAS 102:6273-6278, 2005) and in US Patent Publication Nos. US20070048776 and US20110124100 and International Patent Publication No. WO2007025008, each of which is herein incorporated by reference in its entirety.

“Translational enhancer polynucleotides” or “translation enhancer polynucleotide sequences” are polynucleotides which include one or more of the specific TEE exemplified herein and/or disclosed in the art (see e.g., U.S. Pat. No. 6,310,197, U.S. Pat. No. 6,849,405, U.S. Pat. No. 7,456,273, U.S. Pat. No. 7,183,395, US20090226470, US20070048776, US20110124100, US20090093049, US20130177581, WO2009075886, WO2007025008, WO2012009644, WO2001055371 WO1999024595, and EP2610341A1 and EP2610340A1; each of which is herein incorporated by reference in its entirety) or their variants, homologs or functional derivatives. One or multiple copies of a specific TEE can be present in the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA. The TEEs in the translational enhancer polynucleotides can be organized in one or more sequence segments. A sequence segment can harbor one or more of the specific TEEs exemplified herein, with each TEE being present in one or more copies. When multiple sequence segments are present in a translational enhancer polynucleotide, they can be homogenous or heterogeneous. Thus, the multiple sequence segments in a translational enhancer polynucleotide can harbor identical or different types of the specific TEEs exemplified herein, identical or different number of copies of each of the specific TEEs, and/or identical or different organization of the TEEs within each sequence segment.

In one embodiment, the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA may include at least one TEE that is described in International Patent Publication No. WO1999024595, WO2012009644, WO2009075886, WO2007025008, WO1999024595, European Patent Publication No. EP2610341A1 and EP2610340A1, U.S. Pat. No. 6,310,197, U.S. Pat. No. 6,849,405, U.S. Pat. No. 7,456,273, U.S. Pat. No. 7,183,395, US Patent Publication No. US20090226470, US20110124100, US20070048776, US20090093049 and US20130177581, each of which is herein incorporated by reference in its entirety. The TEE may be located in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA.

In another embodiment, the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA may include at least one TEE that has at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identity with the TEEs described in US Patent Publication Nos. US20090226470, US20070048776, US20130177581 and US20110124100, International Patent Publication No. WO1999024595, WO2012009644, WO2009075886 and WO2007025008, European Patent Publication No. EP2610341A1 and EP2610340A1, U.S. Pat. No. 6,310,197, U.S. Pat. No. 6,849,405, U.S. Pat. No. 7,456,273, U.S. Pat. No. 7,183,395, each of which is herein incorporated by reference in its entirety.

In one embodiment, the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA may include at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18 at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55 or more than 60 TEE sequences. The TEE sequences in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention may be the same or different TEE sequences. The TEE sequences may be in a pattern such as ABABAB or AABBAABBAABB or ABCABCABC or variants thereof repeated once, twice, or more than three times. In these patterns, each letter, A, B, or C represent a different TEE sequence at the nucleotide level.

In one embodiment, the 5′UTR may include a spacer to separate two TEE sequences. As a non-limiting example, the spacer may be a 15 nucleotide spacer and/or other spacers known in the art. As another non-limiting example, the 5′UTR may include a TEE sequence-spacer module repeated at least once, at least twice, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times and at least 9 times or more than 9 times in the 5′UTR.

In another embodiment, the spacer separating two TEE sequences may include other sequences known in the art which may regulate the translation of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention such as, but not limited to, miR sequences described herein (e.g., miR binding sites and miR seeds). As a non-limiting example, each spacer used to separate two TEE sequences may include a different miR sequence or component of a miR sequence (e.g., miR seed sequence).

In one embodiment, the TEE in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention may include at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99% or more than 99% of the TEE sequences disclosed in US Patent Publication Nos. US20090226470, US20070048776, US20130177581 and US20110124100, International Patent Publication No. WO1999024595, WO2012009644, WO2009075886 and WO2007025008, European Patent Publication No. EP2610341A1 and EP2610340A1, U.S. Pat. No. 6,310,197, U.S. Pat. No. 6,849,405, U.S. Pat. No. 7,456,273, U.S. Pat. No. 7,183,395. In another embodiment, the TEE in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention may include a 5-30 nucleotide fragment, a 5-25 nucleotide fragment, a 5-20 nucleotide fragment, a 5-15 nucleotide fragment, a 5-10 nucleotide fragment of the TEE sequences disclosed in US Patent Publication Nos. US20090226470, US20070048776, US20130177581 and US20110124100, International Patent Publication No. WO1999024595, WO2012009644, WO2009075886 and WO2007025008, European Patent Publication No. EP2610341A1 and EP2610340A1, U.S. Pat. No. 6,310,197, U.S. Pat. No. 6,849,405, U.S. Pat. No. 7,456,273, U.S. Pat. No. 7,183,395; each of which are herein incorporated by reference in their entirety.

In one embodiment, the TEE in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention may include at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99% or more than 99% of the TEE sequences disclosed in Chappell et al. (Proc. Natl. Acad. Sci. USA 101:9590-9594, 2004) and Zhou et al. (PNAS 102:6273-6278, 2005), in Supplemental Table 1 and in Supplemental Table 2 disclosed by Wellensiek et al (Genome-wide profiling of human cap-independent translation-enhancing elements, Nature Methods, 2013; DOI:10.1038/NMETH.2522); each of which is herein incorporated by reference in its entirety. In another embodiment, the TEE in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention may include a 5-30 nucleotide fragment, a 5-25 nucleotide fragment, a 5-20 nucleotide fragment, a 5-15 nucleotide fragment, a 5-10 nucleotide fragment of the TEE sequences disclosed in Chappell et al. (Proc. Natl. Acad. Sci. USA 101:9590-9594, 2004) and Zhou et al. (PNAS 102:6273-6278, 2005), in Supplemental Table 1 and in Supplemental Table 2 disclosed by Wellensiek et al (Genome-wide profiling of human cap-independent translation-enhancing elements, Nature Methods, 2013; DOI:10.1038/NMETH.2522); each of which is herein incorporated by reference in its entirety.

In one embodiment, the TEE used in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention is an IRES sequence such as, but not limited to, those described in U.S. Pat. No. 7,468,275 and International Patent Publication No. WO2001055369, each of which is herein incorporated by reference in its entirety.

In one embodiment, the TEEs used in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention may be identified by the methods described in US Patent Publication No. US20070048776 and US20110124100 and International Patent Publication Nos. WO2007025008 and WO2012009644, each of which is herein incorporated by reference in its entirety.

In another embodiment, the TEEs used in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention may be a transcription regulatory element described in U.S. Pat. No. 7,456,273 and U.S. Pat. No. 7,183,395, US Patent Publication No. US20090093049, and International Publication No. WO2001055371, each of which is herein incorporated by reference in their entirety. The transcription regulatory elements may be identified by methods known in the art, such as, but not limited to, the methods described in U.S. Pat. No. 7,456,273 and U.S. Pat. No. 7,183,395, US Patent Publication No. US20090093049, and International Publication No. WO2001055371, each of which is herein incorporated by reference in their entirety.

In yet another embodiment, the TEE used in the 5′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention is an oligonucleotide or portion thereof as described in U.S. Pat. No. 7,456,273 and U.S. Pat. No. 7,183,395, US Patent Publication No. US20090093049, and International Publication No. WO2001055371, each of which is herein incorporated by reference in their entirety.

The 5′ UTR comprising at least one TEE described herein may be incorporated in a monocistronic sequence such as, but not limited to, a vector system or a nucleic acid vector. As a non-limiting example, the vector systems and nucleic acid vectors may include those described in U.S. Pat. No. 7,456,273 and U.S. Pat. No. 7,183,395, US Patent Publication No. US20070048776, US20090093049 and US20110124100 and International Patent Publication Nos. WO2007025008 and WO2001055371, each of which is herein incorporated by reference in its entirety.

In one embodiment, the TEEs described herein may be located in the 5′UTR and/or the 3′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA. The TEEs located in the 3′UTR may be the same and/or different than the TEEs located in and/or described for incorporation in the 5′UTR.

In one embodiment, the 3′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA may include at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18 at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55 or more than 60 TEE sequences. The TEE sequences in the 3′UTR of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention may be the same or different TEE sequences. The TEE sequences may be in a pattern such as ABABAB or AABBAABBAABB or ABCABCABC or variants thereof repeated once, twice, or more than three times. In these patterns, each letter, A, B, or C represent a different TEE sequence at the nucleotide level.

In one embodiment, the 3′UTR may include a spacer to separate two TEE sequences. As a non-limiting example, the spacer may be a 15 nucleotide spacer and/or other spacers known in the art. As another non-limiting example, the 3′UTR may include a TEE sequence-spacer module repeated at least once, at least twice, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times and at least 9 times or more than 9 times in the 3′UTR.

In another embodiment, the spacer separating two TEE sequences may include other sequences known in the art which may regulate the translation of the signal-sensor polynucleotides, primary constructs, modified nucleic acids and/or mmRNA of the present invention such as, but not limited to, miR sequences described herein (e.g., miR binding sites and miR seeds). As a non-limiting example, each spacer used to separate two TEE sequences may include a different miR sequence or component of a miR sequence (e.g., miR seed sequence).

In one embodiment, the incorporation of a miR sequence and/or a TEE sequence changes the shape of the stem loop region which may increase and/or decrease translation. (see e.g, Kedde et al. A Pumilio-induced RNA structure switch in p27-3′UTR controls miR-221 and miR-22 accessibility. Nature Cell Biology. 2010, herein incorporated by reference in its entirety).

In one embodiment, the 5′UTR may comprise at least one microRNA sequence. The microRNA sequence may be, but is not limited to, a 19 or 22 nucleotide sequence and/or a microRNA sequence without the seed.

In one embodiment the microRNA sequence in the 5′UTR may be used to stabilize the nucleic acid and/or mRNA described herein.

In another embodiment, a microRNA sequence in the 5′UTR may be used to decrease the accessibility of the site of translation initiation such as, but not limited to a start codon. Matsuda et al (PLoS One. 2010 11(5):e15057; herein incorporated by reference in its entirety) used antisense locked nucleic acid (LNA) oligonucleotides and exon-junction complexes (EJCs) around a start codon (−4 to +37 where the A of the AUG codons is +1) in order to decrease the accessibility to the first start codon (AUG). Matsuda showed that altering the sequence around the start codon with an LNA or EJC the efficiency, length and structural stability of the nucleic acid or mRNA is affected. The signal-sensor polynucleotides of the present invention may comprise a microRNA sequence, instead of the LNA or EJC sequence described by Matsuda et al, near the site of translation initiation in order to decrease the accessibility to the site of translation initiation. The site of translation initiation may be prior to, after or within the microRNA sequence. As a non-limiting example, the site of translation initiation may be located within a microRNA sequence such as a seed sequence or binding site. As another non-limiting example, the site of translation initiation may be located within a miR-122 sequence such as the seed sequence or the mir-122 binding site.

In one embodiment, the nucleic acids or mRNA of the present invention comprises at least one microRNA sequence in a region of the nucleic acid or mRNA which may interact with a RNA binding protein.

RNA Motifs for RNA Binding Proteins (RBPs)

RNA binding proteins (RBPs) can regulate numerous aspects of co- and post-transcription gene expression such as, but not limited to, RNA splicing, localization, translation, turnover, polyadenylation, capping, modification, export and localization. RNA-binding domains (RBDs), such as, but not limited to, RNA recognition motif (RR) and hnRNP K-homology (KH) domains, typically regulate the sequence association between RBPs and their RNA targets (Ray et al. Nature 2013. 499:172-177; herein incorporated by reference in its entirety). In one embodiment, the canonical RBDs can bind short RNA sequences. In another embodiment, the canonical RBDs can recognize structure RNAs.

In one embodiment, the nucleic acids and/or mRNA may comprise at least one RNA-binding motif such as, but not limited to a RNA-binding domain (RBD).

In one embodiment, the RBD may be any of the RBDs, fragments or variants thereof descried by Ray et al. (Nature 2013. 499:172-177; herein incorporated by reference in its entirety).

In one embodiment, the nucleic acids or mRNA of the present invention may comprise a sequence for at least one RNA-binding domain (RBDs). When the nucleic acids or mRNA of the present invention comprise more than one RBD, the RBDs do not need to be from the same species or even the same structural class.

In one embodiment, at least one flanking region (e.g., the 5′UTR and/or the 3′UTR) may comprise at least one RBD. In another embodiment, the first flanking region and the second flanking region may both comprise at least one RBD. The RBD may be the same or each of the RBDs may have at least 60% sequence identity to the other RBD. As a non-limiting example, at least on RBD may be located before, after and/or within the 3′UTR of the nucleic acid or mRNA of the present invention. As another non-limiting example, at least one RBD may be located before or within the first 300 nucleosides of the 3′UTR.

In another embodiment, the nucleic acids and/or mRNA of the present invention may comprise at least one RBD in the first region of linked nucleosides. The RBD may be located before, after or within a coding region (e.g., the ORF).

In yet another embodiment, the first region of linked nucleosides and/or at least one flanking region may comprise at least on RBD. As a non-limiting example, the first region of linked nucleosides may comprise a RBD related to splicing factors and at least one flanking region may comprise a RBD for stability and/or translation factors.

In one embodiment, the nucleic acids and/or mRNA of the present invention may comprise at least one RBD located in a coding and/or non-coding region of the nucleic acids and/or mRNA.

In one embodiment, at least one RBD may be incorporated into at least one flanking region to increase the stability of the nucleic acid and/or mRNA of the present invention.

In one embodiment, a microRNA sequence in a RNA binding protein motif may be used to decrease the accessibility of the site of translation initiation such as, but not limited to a start codon. The signal-sensor polynucleotides of the present invention may comprise a microRNA sequence, instead of the LNA or EJC sequence described by Matsuda et al, near the site of translation initiation in order to decrease the accessibility to the site of translation initiation. The site of translation initiation may be prior to, after or within the microRNA sequence. As a non-limiting example, the site of translation initiation may be located within a microRNA sequence such as a seed sequence or binding site. As another non-limiting example, the site of translation initiation may be located within a miR-122 sequence such as the seed sequence or the mir-122 binding site.

In another embodiment, an antisense locked nucleic acid (LNA) oligonucleotides and exon-junction complexes (EJCs) may be used in the RNA binding protein motif. The LNA and EJCs may be used around a start codon (−4 to +37 where the A of the AUG codons is +1) in order to decrease the accessibility to the first start codon (AUG).

3′ UTR and the AU Rich Elements

3′UTRs are known to have stretches of Adenosines and Uridines embedded in them. These AU rich signatures are particularly prevalent in genes with high rates of turnover. Based on their sequence features and functional properties, the AU rich elements (AREs) can be separated into three classes (Chen et al, 1995): Class I AREs contain several dispersed copies of an AUUUA motif within U-rich regions. C-Myc and MyoD contain class I AREs. Class II AREs possess two or more overlapping UUAUUUA(U/A)(U/A) nonamers. Molecules containing this type of AREs include GM-CSF and TNF-a. Class III ARES are less well defined. These U rich regions do not contain an AUUUA motif c-Jun and Myogenin are two well-studied examples of this class. Most proteins binding to the AREs are known to destabilize the messenger, whereas members of the ELAV family, most notably HuR, have been documented to increase the stability of mRNA. HuR binds to AREs of all the three classes. Engineering the HuR specific binding sites into the 3′ UTR of nucleic acid molecules will lead to HuR binding and thus, stabilization of the message in vivo.

Introduction, removal or modification of 3′ UTR AU rich elements (AREs) can be used to modulate the stability of signal-sensor polynucleotides, primary constructs or mmRNA of the invention. When engineering specific polynucleotides, primary constructs or mmRNA, one or more copies of an ARE can be introduced to make polynucleotides, primary constructs or mmRNA of the invention less stable and thereby curtail translation and decrease production of the resultant protein. Likewise, AREs can be identified and removed or mutated to increase the intracellular stability and thus increase translation and production of the resultant protein. Transfection experiments can be conducted in relevant cell lines, using signal-sensor polynucleotides, primary constructs or mmRNA of the invention and protein production can be assayed at various time points post-transfection. For example, cells can be transfected with different ARE-engineering molecules and by using an ELISA kit to the relevant protein and assaying protein produced at 6 hr, 12 hr, 24 hr, 48 hr, and 7 days post-transfection.

3′ UTR and Triple Helices

In one embodiment, signal-sequence polynucleotides of the present invention may include a triple helix on the 3′ end of the signal-sequence polynucleotides. The 3′ end of the nucleic acids of the present invention may include a triple helix alone or in combination with a Poly-A tail.

In one embodiment, the signal-sequence polynucleotides of the present invention may comprise at least a first and a second U-rich region, a conserved stem loop region between the first and second region and an A-rich region. The first and second U-rich region and the A-rich region may associate to form a triple helix on the 3′ end of the nucleic acid. This triple helix may stabilize the nucleic acid, enhance the translational efficiency of the nucleic acid and/or protect the 3′ end from degradation. Exemplary triple helices include, but are not limited to, the triple helix sequence of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), MEN-β and polyadenylated nuclear (PAN) RNA (See Wilusz et al., Genes & Development 2012 26:2392-2407; herein incorporated by reference in its entirety). In one embodiment, the 3′ end of the modified nucleic acids, enhanced modified RNA or ribonucleic acids of the present invention comprises a first U-rich region comprising TTTTTCTTTT (SEQ ID NO: 1), a second U-rich region comprising TTTTGCTTTTT (SEQ ID NO: 2) or TTTTGCTTTT (SEQ ID NO: 3), an A-rich region comprising AAAAAGCAAAA (SEQ ID NO: 4). In another embodiment, the 3′ end of the nucleic acids of the present invention comprises a triple helix formation structure comprising a first U-rich region, a conserved region, a second U-rich region and an A-rich region.

In one embodiment, the triple helix may be formed from the cleavage of a MALAT1 sequence prior to the cloverleaf structure. While not meaning to be bound by theory, MALAT1 is a long non-coding RNA which, when cleaved, forms a triple helix and a tRNA-like cloverleaf structure. The MALAT1 transcript then localizes to nuclear speckles and the tRNA-like cloverleaf localizes to the cytoplasm (Wilusz et al. Cell 2008 135(5): 919-932; herein incorporated by reference in its entirety).

As a non-limiting example, the terminal end of the nucleic acid of the present invention comprising the MALAT1 sequence can then form a triple helix structure, after RNaseP cleavage from the cloverleaf structure, which stabilizes the nucleic acid (Peart et al. Non-mRNA 3′ end formation: how the other half lives; WIREs RNA 2013; herein incorporated by reference in its entirety).

In one embodiment, the signal-sequence polynucleotides described herein comprise a MALAT1 sequence. In another embodiment, the signal-sequence polynucleotides may be polyadenylated. In yet another embodiment, the signal-sequence polynucleotides is not polyadenylated but has an increased resistance to degradation compared to unmodified nucleic acids or mRNA.

In one embodiment, the signal-sequence polynucleotides of the present invention may comprise a MALAT1 sequence in the second flanking region (e.g., the 3′UTR). As a non-limiting example, the MALAT1 sequence may be human or mouse.

In another embodiment, the cloverleaf structure of the MALAT1 sequence may also undergo processing by RNaseZ and CCA adding enzyme to form a tRNA-like structure called mascRNA (MALAT1-associated small cytoplasmic RNA). As a non-limiting example, the mascRNA may encode a protein or a fragment thereof and/or may comprise a microRNA sequence. The mascRNA may comprise at least one chemical modification described herein.

Stem Loop

In one embodiment, the nucleic acids of the present invention may include a stem loop such as, but not limited to, a histone stem loop. The stem loop may be a nucleotide sequence that is about 25 or about 26 nucleotides in length such as, but not limited to, SEQ ID NOs: 7-17 as described in International Patent Publication No. WO2013103659, herein incorporated by reference in its entirety. The histone stem loop may be located 3′ relative to the coding region (e.g., at the 3′ terminus of the coding region). As a non-limiting example, the stem loop may be located at the 3′ end of a nucleic acid described herein.

In one embodiment, the stem loop may be located in the second terminal region. As a non-limiting example, the stem loop may be located within an untranslated region (e.g., 3′UTR) in the second terminal region.

In one embodiment, the nucleic acid such as, but not limited to mRNA, which comprises the histone stem loop may be stabilized by the addition of at least one chain terminating nucleoside. Not wishing to be bound by theory, the addition of at least one chain terminating nucleoside may slow the degradation of a nucleic acid and thus can increase the half-life of the nucleic acid.

In one embodiment, the chain terminating nucleoside may be, but is not limited to, those described in International Patent Publication No. WO2013103659, herein incorporated by reference in its entirety. In another embodiment, the chain terminating nucleosides which may be used with the present invention includes, but is not limited to, 3′-deoxyadenosine (cordycepin), 3′-deoxyuridine, 3′-deoxycytosine, 3′-deoxyguanosine, 3′-deoxythymine, 2′,3′-dideoxynucleosides, such as 2′,3′-dideoxyadenosine, 2′,3′-dideoxyuridine, 2′,3′-dideoxycytosine, 2′,3′-dideoxyguanosine, 2′,3′-dideoxythymine, a 2′-deoxynucleoside, or a —O-methylnucleoside.

In another embodiment, the nucleic acid such as, but not limited to mRNA, which comprises the histone stem loop may be stabilized by a modification to the 3′region of the nucleic acid that can prevent and/or inhibit the addition of oligio(U) (see e.g., International Patent Publication No. WO2013103659, herein incorporated by reference in its entirety).

In yet another embodiment, the nucleic acid such as, but not limited to mRNA, which comprises the histone stem loop may be stabilized by the addition of an oligonucleotide that terminates in a 3′-deoxynucleoside, 2′,3′-dideoxynucleoside 3″-0-methylnucleosides, 3′-0-ethylnucleosides, 3′-arabinosides, and other modified nucleosides known in the art and/or described herein.

In one embodiment, the nucleic acids of the present invention may include a histone stem loop, a polyA tail sequence and/or a 5′cap structure. The histone stem loop may be before and/or after the polyA tail sequence. The nucleic acids comprising the histone stem loop and a polyA tail sequence may include a chain terminating nucleoside described herein.

In another embodiment, the nucleic acids of the present invention may include a histone stem loop and a 5′cap structure. The 5′cap structure may include, but is not limited to, those described herein and/or known in the art.

In one embodiment, the conserved stem loop region may comprise a miR sequence described herein. As a non-limiting example, the stem loop region may comprise the seed sequence of a miR sequence described herein. In another non-limiting example, the stem loop region may comprise a miR-122 seed sequence.

In another embodiment, the conserved stem loop region may comprise a miR sequence described herein and may also include a TEE sequence.

In one embodiment, the incorporation of a miR sequence and/or a TEE sequence changes the shape of the stem loop region which may increase and/or decrease translation. (see e.g, Kedde et al. A Pumilio-induced RNA structure switch in p27-3′UTR controls miR-221 and miR-22 accessibility. Nature Cell Biology. 2010, herein incorporated by reference in its entirety).

5′ Capping

The 5′ cap structure of an mRNA is involved in nuclear export, increasing mRNA stability and binds the mRNA Cap Binding Protein (CBP), which is responsible for mRNA stability in the cell and translation competency through the association of CBP with poly(A) binding protein to form the mature cyclic mRNA species. The cap further assists the removal of 5′ proximal introns removal during mRNA splicing.

Endogenous mRNA molecules may be 5′-end capped generating a 5′-ppp-5′-triphosphate linkage between a terminal guanosine cap residue and the 5′-terminal transcribed sense nucleotide of the mRNA molecule. This 5′-guanylate cap may then be methylated to generate an N7-methyl-guanylate residue. The ribose sugars of the terminal and/or anteterminal transcribed nucleotides of the 5′ end of the mRNA may optionally also be 2′-O-methylated. 5′-decapping through hydrolysis and cleavage of the guanylate cap structure may target a nucleic acid molecule, such as an mRNA molecule, for degradation.

Modifications to the signal-sensor polynucleotides, primary constructs, and mmRNA of the present invention may generate a non-hydrolyzable cap structure preventing decapping and thus increasing mRNA half-life. Because cap structure hydrolysis requires cleavage of 5′-ppp-5′ phosphorodiester linkages, modified nucleotides may be used during the capping reaction. For example, a Vaccinia Capping Enzyme from New England Biolabs (Ipswich, Mass.) may be used with α-thio-guanosine nucleotides according to the manufacturer's instructions to create a phosphorothioate linkage in the 5′-ppp-5′ cap. Additional modified guanosine nucleotides may be used such as α-methyl-phosphonate and seleno-phosphate nucleotides.

Additional modifications include, but are not limited to, 2′-O-methylation of the ribose sugars of 5′-terminal and/or 5′-anteterminal nucleotides of the mRNA (as mentioned above) on the 2′-hydroxyl group of the sugar ring. Multiple distinct 5′-cap structures can be used to generate the 5′-cap of a nucleic acid molecule, such as an mRNA molecule.

Cap analogs, which herein are also referred to as synthetic cap analogs, chemical caps, chemical cap analogs, or structural or functional cap analogs, differ from natural (i.e. endogenous, wild-type or physiological) 5′-caps in their chemical structure, while retaining cap function. Cap analogs may be chemically (i.e. non-enzymatically) or enzymatically synthesized and/linked to a nucleic acid molecule.

For example, the Anti-Reverse Cap Analog (ARCA) cap contains two guanines linked by a 5′-5′-triphosphate group, wherein one guanine contains an N7 methyl group as well as a 3′-O-methyl group (i.e., N7,3′-O-dimethyl-guanosine-5′-triphosphate-5′-guanosine (m7G-3′mppp-G; which may equivalently be designated 3′ 0-Me-m7G(5′)ppp(5′)G). The 3′-O atom of the other, unmodified, guanine becomes linked to the 5′-terminal nucleotide of the capped nucleic acid molecule (e.g. an mRNA or mmRNA). The N7- and 3′-O-methylated guanine provides the terminal moiety of the capped nucleic acid molecule (e.g. mRNA or mmRNA).

Another exemplary cap is mCAP, which is similar to ARCA but has a 2′-O-methyl group on guanosine (i.e., N7,2′-O-dimethyl-guanosine-5′-triphosphate-5′-guanosine, m7Gm-ppp-G).

While cap analogs allow for the concomitant capping of a nucleic acid molecule in an in vitro transcription reaction, up to 20% of transcripts remain uncapped. This, as well as the structural differences of a cap analog from an endogenous 5′-cap structures of nucleic acids produced by the endogenous, cellular transcription machinery, may lead to reduced translational competency and reduced cellular stability.

Signal-sensor polynucleotides, primary constructs and mmRNA of the invention may also be capped post-transcriptionally, using enzymes, in order to generate more authentic 5′-cap structures. As used herein, the phrase “more authentic” refers to a feature that closely mirrors or mimics, either structurally or functionally, an endogenous or wild type feature. That is, a “more authentic” feature is better representative of an endogenous, wild-type, natural or physiological cellular function and/or structure as compared to synthetic features or analogs, etc., of the prior art, or which outperforms the corresponding endogenous, wild-type, natural or physiological feature in one or more respects. Non-limiting examples of more authentic 5′cap structures of the present invention are those which, among other things, have enhanced binding of cap binding proteins, increased half life, reduced susceptibility to 5′ endonucleases and/or reduced 5′decapping, as compared to synthetic 5′cap structures known in the art (or to a wild-type, natural or physiological 5′cap structure). For example, recombinant Vaccinia Virus Capping Enzyme and recombinant 2′-O-methyltransferase enzyme can create a canonical 5′-5′-triphosphate linkage between the 5′-terminal nucleotide of an mRNA and a guanine cap nucleotide wherein the cap guanine contains an N7 methylation and the 5′-terminal nucleotide of the mRNA contains a 2′-O-methyl. Such a structure is termed the Cap1 structure. This cap results in a higher translational-competency and cellular stability and a reduced activation of cellular pro-inflammatory cytokines, as compared, e.g., to other 5′cap analog structures known in the art. Cap structures include 7mG(5′)ppp(5′)N,pN2p (cap 0), 7mG(5′)ppp(5′)NlmpNp (cap 1), and 7mG(5′)-ppp(5′)NlmpN2mp (cap 2).

Because the signal-sensor polynucleotides, primary constructs or mmRNA may be capped post-transcriptionally, and because this process is more efficient, nearly 100% of the signal-sensor polynucleotides, primary constructs or mmRNA may be capped. This is in contrast to ˜80% when a cap analog is linked to an mRNA in the course of an in vitro transcription reaction.

According to the present invention, 5′ terminal caps may include endogenous caps or cap analogs. According to the present invention, a 5′ terminal cap may comprise a guanine analog. Useful guanine analogs include inosine, N1-methyl-guanosine, 2′fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

Viral Sequences

Additional viral sequences such as, but not limited to, the translation enhancer sequence of the barley yellow dwarf virus (BYDV-PAV) can be engineered and inserted in the 3′ UTR of the signal-sensor polynucleotides, primary constructs or mmRNA of the invention and can stimulate the translation of the construct in vitro and in vivo. Transfection experiments can be conducted in relevant cell lines at and protein production can be assayed by ELISA at 12 hr, 24 hr, 48 hr, 72 hr and day 7 post-transfection.

IRES Sequences

Further, provided are signal-sensor polynucleotides, primary constructs or mmRNA which may contain an internal ribosome entry site (IRES). First identified as a feature Picorna virus RNA, IRES plays an important role in initiating protein synthesis in absence of the 5′ cap structure. An IRES may act as the sole ribosome binding site, or may serve as one of multiple ribosome binding sites of an mRNA. signal-sensor polynucleotides, primary constructs or mmRNA containing more than one functional ribosome binding site may encode several oncology-related peptides or oncology-related polypeptides that are translated independently by the ribosomes (“multicistronic nucleic acid molecules”). When signal-sensor polynucleotides, primary constructs or mmRNA are provided with an IRES, further optionally provided is a second translatable region. Examples of IRES sequences that can be used according to the invention include without limitation, those from picornaviruses (e.g. FMDV), pest viruses (CFFV), polio viruses (PV), encephalomyocarditis viruses (ECMV), foot-and-mouth disease viruses (FMDV), hepatitis C viruses (HCV), classical swine fever viruses (CSFV), murine leukemia virus (MLV), simian immune deficiency viruses (SIV) or cricket paralysis viruses (CrPV).

Poly-A Tails

During RNA processing, a long chain of adenine nucleotides (poly-A tail) may be added to a polynucleotide such as an mRNA molecule in order to increase stability. Immediately after transcription, the 3′ end of the transcript may be cleaved to free a 3′ hydroxyl. Then poly-A polymerase adds a chain of adenine nucleotides to the RNA. The process, called polyadenylation, adds a poly-A tail that can be between 100 and 250 residues long.

It has been discovered that unique poly-A tail lengths provide certain advantages to the signal-sensor polynucleotides, primary constructs or mmRNA of the present invention.

Generally, the length of a poly-A tail of the present invention is greater than 30 nucleotides in length. In another embodiment, the poly-A tail is greater than 35 nucleotides in length (e.g., at least or greater than about 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,500, and 3,000 nucleotides). In some embodiments, the signal-sensor polynucleotides, primary construct, or mmRNA includes from about 30 to about 3,000 nucleotides (e.g., from 30 to 50, from 30 to 100, from 30 to 250, from 30 to 500, from 30 to 750, from 30 to 1,000, from 30 to 1,500, from 30 to 2,000, from 30 to 2,500, from 50 to 100, from 50 to 250, from 50 to 500, from 50 to 750, from 50 to 1,000, from 50 to 1,500, from 50 to 2,000, from 50 to 2,500, from 50 to 3,000, from 100 to 500, from 100 to 750, from 100 to 1,000, from 100 to 1,500, from 100 to 2,000, from 100 to 2,500, from 100 to 3,000, from 500 to 750, from 500 to 1,000, from 500 to 1,500, from 500 to 2,000, from 500 to 2,500, from 500 to 3,000, from 1,000 to 1,500, from 1,000 to 2,000, from 1,000 to 2,500, from 1,000 to 3,000, from 1,500 to 2,000, from 1,500 to 2,500, from 1,500 to 3,000, from 2,000 to 3,000, from 2,000 to 2,500, and from 2,500 to 3,000).

In one embodiment, the poly-A tail is designed relative to the length of the overall signal-sensor polynucleotides, primary constructs or mmRNA. This design may be based on the length of the coding region, the length of a particular feature or region (such as the first or flanking regions), or based on the length of the ultimate product expressed from the polynucleotides, primary constructs or mmRNA.

In this context the poly-A tail may be 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100% greater in length than the signal-sensor polynucleotides, primary constructs or mmRNA or feature thereof. The poly-A tail may also be designed as a fraction of polynucleotides, primary constructs or mmRNA to which it belongs. In this context, the poly-A tail may be 10, 20, 30, 40, 50, 60, 70, 80, or 90% or more of the total length of the construct or the total length of the construct minus the poly-A tail.

In one embodiment, engineered binding sites and/or conjugation of signal-sensor polynucleotides, primary constructs or mmRNA for Poly-A binding protein may be used to enhance expression. The engineered binding sites may be sensor sequences which can operate as binding sites for ligands of the local microenvironment of the nucleic acids and/or mRNA. As a non-limiting example, the nucleic acids and/or mRNA may comprise at least one engineered binding site to alter the binding affinity of Poly-A binding protein (PABP) and analogs thereof. The incorporation of at least one engineered binding site may increase the binding affinity of the PABP and analogs thereof.

Additionally, multiple distinct signal-sensor polynucleotides, primary constructs or mmRNA may be linked together to the PABP (Poly-A binding protein) through the 3′-end using modified nucleotides at the 3′-terminus of the poly-A tail. Transfection experiments can be conducted in relevant cell lines and protein production can be assayed by ELISA at 12 hr, 24 hr, 48 hr, 72 hr and day 7 post-transfection. As a non-limiting example, the transfection experiments may be used to evaluate the effect on PABP or analogs thereof binding affinity as a result of the addition of at least one engineered binding site.

In one embodiment, the signal-sensor polynucleotides and primary constructs of the present invention are designed to include a polyA-G Quartet. The G-quartet is a cyclic hydrogen bonded array of four guanine nucleotides that can be formed by G-rich sequences in both DNA and RNA. In this embodiment, the G-quartet is incorporated at the end of the poly-A tail. The resultant mmRNA construct is assayed for stability, protein production and other parameters including half-life at various time points. It has been discovered that the polyA-G quartet results in protein production equivalent to at least 75% of that seen using a poly-A tail of 120 nucleotides alone.

In one embodiment, the nucleic acids or mRNA of the present invention may comprise a polyA tail and may be stabilized by the addition of a chain terminating nucleoside. The nucleic acids and/or mRNA with a polyA tail may further comprise a 5′ cap structure.

In another embodiment, the nucleic acids or mRNA of the present invention may comprise a polyA-G Quartet. The nucleic acids and/or mRNA with a polyA-G Quartet may further comprise a 5′ cap structure.

In one embodiment, the chain terminating nucleoside which may be used to stabilize the nucleic acid or mRNA comprising a polyA tail or polyA-G Quartet may be, but is not limited to, those described in International Patent Publication No. WO2013103659, herein incorporated by reference in its entirety. In another embodiment, the chain terminating nucleosides which may be used with the present invention includes, but is not limited to, 3′-deoxyadenosine (cordycepin), 3′-deoxyuridine, 3′-deoxycytosine, 3′-deoxyguanosine, 3′-deoxythymine, 2′,3′-dideoxynucleosides, such as 2′,3′-dideoxyadenosine, 2′,3′-dideoxyuridine, 2′,3′-dideoxycytosine, 2′,3′-dideoxyguanosine, 2′,3′-dideoxythymine, a 2′-deoxynucleoside, or a —O-methylnucleoside.

In another embodiment, the nucleic acid such as, but not limited to mRNA, which comprise a polyA tail or a polyA-G Quartet may be stabilized by a modification to the 3′region of the nucleic acid that can prevent and/or inhibit the addition of oligio(U) (see e.g., International Patent Publication No. WO2013103659, herein incorporated by reference in its entirety).

In yet another embodiment, the nucleic acid such as, but not limited to mRNA, which comprise a polyA tail or a polyA-G Quartet may be stabilized by the addition of an oligonucleotide that terminates in a 3′-deoxynucleoside, 2′,3′-dideoxynucleoside 3′-0-methylnucleosides, 3′-0-ethylnucleosides, 3′-arabinosides, and other modified nucleosides known in the art and/or described herein.

Quantification

In one embodiment, the signal-sensor polynucleotides, primary constructs or mmRNA of the present invention may be quantified in exosomes derived from one or more bodily fluid. As used herein “bodily fluids” include peripheral blood, serum, plasma, ascites, urine, cerebrospinal fluid (CSF), sputum, saliva, bone marrow, synovial fluid, aqueous humor, amniotic fluid, cerumen, breast milk, broncheoalveolar lavage fluid, semen, prostatic fluid, cowper's fluid or pre-ejaculatory fluid, sweat, fecal matter, hair, tears, cyst fluid, pleural and peritoneal fluid, pericardial fluid, lymph, chyme, chyle, bile, interstitial fluid, menses, pus, sebum, vomit, vaginal secretions, mucosal secretion, stool water, pancreatic juice, lavage fluids from sinus cavities, bronchopulmonary aspirates, blastocyl cavity fluid, and umbilical cord blood. Alternatively, exosomes may be retrieved from an organ selected from the group consisting of lung, heart, pancreas, stomach, intestine, bladder, kidney, ovary, testis, skin, colon, breast, prostate, brain, esophagus, liver, and placenta.

In the quantification method, a sample of not more than 2 mL is obtained from the subject and the exosomes isolated by size exclusion chromatography, density gradient centrifugation, differential centrifugation, nanomembrane ultrafiltration, immunoabsorbent capture, affinity purification, microfluidic separation, or combinations thereof. In the analysis, the level or concentration of signal-sensor polynucleotides, primary construct or mmRNA may be an expression level, presence, absence, truncation or alteration of the administered construct. It is advantageous to correlate the level with one or more clinical phenotypes or with an assay for a human disease biomarker. The assay may be performed using construct specific probes, cytometry, qRT-PCR, real-time PCR, PCR, flow cytometry, electrophoresis, mass spectrometry, or combinations thereof while the exosomes may be isolated using immunohistochemical methods such as enzyme linked immunosorbent assay (ELISA) methods. Exosomes may also be isolated by size exclusion chromatography, density gradient centrifugation, differential centrifugation, nanomembrane ultrafiltration, immunoabsorbent capture, affinity purification, microfluidic separation, or combinations thereof.

These methods afford the investigator the ability to monitor, in real time, the level of signal-sensor polynucleotides, primary constructs or mmRNA remaining or delivered. This is possible because the polynucleotides, primary constructs or mmRNA of the present invention differ from the endogenous forms due to the structural and/or chemical modifications.

II. DESIGN AND SYNTHESIS OF SIGNAL-SENSOR POLYNUCLEOTIDES

Signal-sensor polynucleotides, primary constructs or mmRNA for use in accordance with the invention may be prepared according to any available technique including, but not limited to chemical synthesis, enzymatic synthesis, which is generally termed in vitro transcription (IVT) or enzymatic or chemical cleavage of a longer precursor, etc. Methods of synthesizing RNAs are known in the art (see, e.g., Gait, M. J. (ed.) Oligonucleotide synthesis: a practical approach, Oxford [Oxfordshire], Washington, D.C.: IRL Press, 1984; and Herdewijn, P. (ed.) Oligonucleotide synthesis: methods and applications, Methods in Molecular Biology, v. 288 (Clifton, N.J.) Totowa, N.J.: Humana Press, 2005; both of which are incorporated herein by reference).

The process of design and synthesis of the signal-sensor primary constructs of the invention generally includes the steps of gene construction, mRNA production (either with or without modifications) and purification. In the enzymatic synthesis method, a target signal-sensor polynucleotide sequence encoding the oncology-related polypeptide of interest is first selected for incorporation into a vector which will be amplified to produce a cDNA template. Optionally, the target signal-sensor polynucleotide sequence and/or any flanking sequences may be codon optimized. The cDNA template is then used to produce mRNA through in vitro transcription (IVT). After production, the mRNA may undergo purification and clean-up processes. The steps of which are provided in more detail below.

Gene Construction

The step of gene construction may include, but is not limited to gene synthesis, vector amplification, plasmid purification, plasmid linearization and clean-up, and cDNA template synthesis and clean-up.

Gene Synthesis

Once an oncology-related polypeptide of interest, or target, is selected for production, a signal-sensor primary construct is designed. Within the primary construct, a first region of linked nucleosides encoding the polypeptide of interest may be constructed using an open reading frame (ORF) of a selected nucleic acid (DNA or RNA) transcript. The ORF may comprise the wild type ORF, an isoform, variant or a fragment thereof. As used herein, an “open reading frame” or “ORF” is meant to refer to a nucleic acid sequence (DNA or RNA) which is capable of encoding an oncology-related polypeptide of interest. ORFs often begin with the start codon, ATG and end with a nonsense or termination codon or signal.

Further, the nucleotide sequence of the first region may be codon optimized. Codon optimization methods are known in the art and may be useful in efforts to achieve one or more of several goals. These goals include to match codon frequencies in target and host organisms to ensure proper folding, bias GC content to increase mRNA stability or reduce secondary structures, minimize tandem repeat codons or base runs that may impair gene construction or expression, customize transcriptional and translational control regions, insert or remove protein trafficking sequences, remove/add post translation modification sites in encoded protein (e.g. glycosylation sites), add, remove or shuffle protein domains, insert or delete restriction sites, modify ribosome binding sites and mRNA degradation sites, to adjust translational rates to allow the various domains of the protein to fold properly, or to reduce or eliminate problem secondary structures within the mRNA. Codon optimization tools, algorithms and services are known in the art, non-limiting examples include services from GeneArt (Life Technologies) and/or DNA2.0 (Menlo Park Calif.). In one embodiment, the ORF sequence is optimized using optimization algorithms. Codon options for each amino acid are given in Table 1.

TABLE 1
Codon Options
Single
Letter
Amino AcidCodeCodon Options
IsoleucineIATT, ATC, ATA
LeucineLCTT, CTC, CTA, CTG,
TTA, TTG
ValineVGTT, GTC, GTA, GTG
PhenylalanineFTTT, TTC
MethionineMATG
CysteineCTGT, TGC
AlanineAGCT, GCC, GCA, GCG
GlycineGGGT, GGC, GGA, GGG
ProlinePCCT, CCC, CCA, CCG
ThreonineTACT, ACC, ACA, ACG
SerineSTCT, TCC, TCA, TCG,
AGT, AGC
TyrosineYTAT, TAC
TryptophanWTGG
GlutamineQCAA, CAG
AsparagineNAAT, AAC
HistidineHCAT, CAC
Glutamic acidEGAA, GAG
Aspartic acidDGAT, GAC
LysineKAAA, AAG
ArginineRCGT, CGC, CGA, CGG,
AGA, AGG
SelenocysteineSecUGA in mRNA in presence
of Selenocystein insertion
element (SECIS)
Stop codonsStopTAA, TAG, TGA

In one embodiment, after a nucleotide sequence has been codon optimized it may be further evaluated for regions containing restriction sites. At least one nucleotide within the restriction site regions may be replaced with another nucleotide in order to remove the restriction site from the sequence but the replacement of nucleotides does alter the amino acid sequence which is encoded by the codon optimized nucleotide sequence.

Features, which may be considered beneficial in some embodiments of the present invention, may be encoded by the signal-sensor primary construct and may flank the ORF as a first or second flanking region. The flanking regions may be incorporated into the signal-sensor primary construct before and/or after optimization of the ORF. It is not required that a signal-sensor primary construct contain both a 5′ and 3′ flanking region. Examples of such features include, but are not limited to, untranslated regions (UTRs), Kozak sequences, an oligo(dT) sequence, and detectable tags and may include multiple cloning sites which may have XbaI recognition.

In some embodiments, a 5′ UTR and/or a 3′ UTR may be provided as flanking regions. Multiple 5′ or 3′ UTRs may be included in the flanking regions and may be the same or of different sequences. Any portion of the flanking regions, including none, may be codon optimized and any may independently contain one or more different structural or chemical modifications, before and/or after codon optimization. Combinations of features may be included in the first and second flanking regions and may be contained within other features. For example, the ORF may be flanked by a 5′ UTR which may contain a strong Kozak translational initiation signal and/or a 3′ UTR which may include an oligo(dT) sequence for templated addition of a poly-A tail.

Tables 2 and 3 provide a listing of exemplary UTRs which may be utilized in the signal-sensor primary construct of the present invention as flanking regions. Shown in Table 2 is a representative listing of a 5′-untranslated region of the invention. Variants of 5′ UTRs may be utilized wherein one or more nucleotides are added or removed to the termini, including A, T, C or G.

TABLE 2
5′-Untranslated Regions
5′ UTRName/SEQ ID
IdentifierDescriptionSequenceNO.
NativeWild type UTRSee wild type sequence
5UTR-001Synthetic UTRGGGAAATAAGAGAGAAAAGAAGAGTAAGA1
AGAAATATAAGAGCCACC
5UTR-002Upstream UTRGGGAGATCAGAGAGAAAAGAAGAGTAAGA2
AGAAATATAAGAGCCACC
5UTR-003Upstream UTRGGAATAAAAGTCTCAACACAACATATACAA3
AACAAACGAATCTCAAGCAATCAAGCATTC
TACTTCTATTGCAGCAATTTAAATCATTTCT
TTTAAAGCAAAAGCAATTTTCTGAAAATTT
TCACCATTTACGAACGATAGCAAC
5UTR-004Upstream UTRGGGAGACAAGCUUGGCAUUCCGGUACUGU4
UGGUAAAGCCACC

Shown in Table 3 is a representative listing of 3′-untranslated regions of the invention. Variants of 3′ UTRs may be utilized wherein one or more nucleotides are added or removed to the termini, including A, T, C or G.

TABLE 3
3′-Untranslated Regions
SEQ
3′ UTRName/ID
IdentifierDescriptionSequenceNO.
3UTR-001CreatineGCGCCTGCCCACCTGCCACCGACTGCTGGAACC5
KinaseCAGCCAGTGGGAGGGCCTGGCCCACCAGAGTCC
TGCTCCCTCACTCCTCGCCCCGCCCCCTGTCCCA
GAGTCCCACCTGGGGGCTCTCTCCACCCTTCTCA
GAGTTCCAGTTTCAACCAGAGTTCCAACCAATG
GGCTCCATCCTCTGGATTCTGGCCAATGAAATAT
CTCCCTGGCAGGGTCCTCTTCTTTTCCCAGAGCT
CCACCCCAACCAGGAGCTCTAGTTAATGGAGAG
CTCCCAGCACACTCGGAGCTTGTGCTTTGTCTCC
ACGCAAAGCGATAAATAAAAGCATTGGTGGCCT
TTGGTCTTTGAATAAAGCCTGAGTAGGAAGTCTA
GA
3UTR-002MyoglobinGCCCCTGCCGCTCCCACCCCCACCCATCTGGGCC6
CCGGGTTCAAGAGAGAGCGGGGTCTGATCTCGT
GTAGCCATATAGAGTTTGCTTCTGAGTGTCTGCT
TTGTTTAGTAGAGGTGGGCAGGAGGAGCTGAGG
GGCTGGGGCTGGGGTGTTGAAGTTGGCTTTGCAT
GCCCAGCGATGCGCCTCCCTGTGGGATGTCATCA
CCCTGGGAACCGGGAGTGGCCCTTGGCTCACTG
TGTTCTGCATGGTTTGGATCTGAATTAATTGTCC
TTTCTTCTAAATCCCAACCGAACTTCTTCCAACC
TCCAAACTGGCTGTAACCCCAAATCCAAGCCATT
AACTACACCTGACAGTAGCAATTGTCTGATTAAT
CACTGGCCCCTTGAAGACAGCAGAATGTCCCTTT
GCAATGAGGAGGAGATCTGGGCTGGGCGGGCCA
GCTGGGGAAGCATTTGACTATCTGGAACTTGTGT
GTGCCTCCTCAGGTATGGCAGTGACTCACCTGGT
TTTAATAAAACAACCTGCAACATCTCATGGTCTT
TGAATAAAGCCTGAGTAGGAAGTCTAGA
3UTR-003α-actinACACACTCCACCTCCAGCACGCGACTTCTCAGG7
ACGACGAATCTTCTCAATGGGGGGGCGGCTGAG
CTCCAGCCACCCCGCAGTCACTTTCTTTGTAACA
ACTTCCGTTGCTGCCATCGTAAACTGACACAGTG
TTTATAACGTGTACATACATTAACTTATTACCTC
ATTTTGTTATTTTTCGAAACAAAGCCCTGTGGAA
GAAAATGGAAAACTTGAAGAAGCATTAAAGTCA
TTCTGTTAAGCTGCGTAAATGGTCTTTGAATAAA
GCCTGAGTAGGAAGTCTAGA
3UTR-004AlbuminCATCACATTTAAAAGCATCTCAGCCTACCATGAG8
AATAAGAGAAAGAAAATGAAGATCAAAAGCTT
ATTCATCTGTTTTTCTTTTTCGTTGGTGTAAAGCC
AACACCCTGTCTAAAAAACATAAATTTCTTTAAT
CATTTTGCCTCTTTTCTCTGTGCTTCAATTAATAA
AAAATGGAAAGAATCTAATAGAGTGGTACAGCA
CTGTTATTTTTCAAAGATGTGTTGCTATCCTGAA
AATTCTGTAGGTTCTGTGGAAGTTCCAGTGTTCT
CTCTTATTCCACTTCGGTAGAGGATTTCTAGTTT
CTTGTGGGCTAATTAAATAAATCATTAATACTCT
TCTAATGGTCTTTGAATAAAGCCTGAGTAGGAA
GTCTAGA
3UTR-005α-globinGCTGCCTTCTGCGGGGCTTGCCTTCTGGCCATGC9
CCTTCTTCTCTCCCTTGCACCTGTACCTCTTGGTC
TTTGAATAAAGCCTGAGTAGGAAGGCGGCCGCT
CGAGCATGCATCTAGA
3UTR-006G-CSFGCCAAGCCCTCCCCATCCCATGTATTTATCTCTA10
TTTAATATTTATGTCTATTTAAGCCTCATATTTAA
AGACAGGGAAGAGCAGAACGGAGCCCCAGGCC
TCTGTGTCCTTCCCTGCATTTCTGAGTTTCATTCT
CCTGCCTGTAGCAGTGAGAAAAAGCTCCTGTCCT
CCCATCCCCTGGACTGGGAGGTAGATAGGTAAA
TACCAAGTATTTATTACTATGACTGCTCCCCAGC
CCTGGCTCTGCAATGGGCACTGGGATGAGCCGC
TGTGAGCCCCTGGTCCTGAGGGTCCCCACCTGGG
ACCCTTGAGAGTATCAGGTCTCCCACGTGGGAG
ACAAGAAATCCCTGTTTAATATTTAAACAGCAGT
GTTCCCCATCTGGGTCCTTGCACCCCTCACTCTG
GCCTCAGCCGACTGCACAGCGGCCCCTGCATCC
CCTTGGCTGTGAGGCCCCTGGACAAGCAGAGGT
GGCCAGAGCTGGGAGGCATGGCCCTGGGGTCCC
ACGAATTTGCTGGGGAATCTCGTTTTTCTTCTTA
AGACTTTTGGGACATGGTTTGACTCCCGAACATC
ACCGACGCGTCTCCTGTTTTTCTGGGTGGCCTCG
GGACACCTGCCCTGCCCCCACGAGGGTCAGGAC
TGTGACTCTTTTTAGGGCCAGGCAGGTGCCTGGA
CATTTGCCTTGCTGGACGGGGACTGGGGATGTG
GGAGGGAGCAGACAGGAGGAATCATGTCAGGC
CTGTGTGTGAAAGGAAGCTCCACTGTCACCCTCC
ACCTCTTCACCCCCCACTCACCAGTGTCCCCTCC
ACTGTCACATTGTAACTGAACTTCAGGATAATAA
AGTGTTTGCCTCCATGGTCTTTGAATAAAGCCTG
AGTAGGAAGGCGGCCGCTCGAGCATGCATCTAGA
3UTR-007Col1a2;ACTCAATCTAAATTAAAAAAGAAAGAAATTTGA11
collagen,AAAAACTTTCTCTTTGCCATTTCTTCTTCTTCTTT
type I, alpha 2TTTAACTGAAAGCTGAATCCTTCCATTTCTTCTG
CACATCTACTTGCTTAAATTGTGGGCAAAAGAG
AAAAAGAAGGATTGATCAGAGCATTGTGCAATA
CAGTTTCATTAACTCCTTCCCCCGCTCCCCCAAA
AATTTGAATTTTTTTTTCAACACTCTTACACCTGT
TATGGAAAATGTCAACCTTTGTAAGAAAACCAA
AATAAAAATTGAAAAATAAAAACCATAAACATT
TGCACCACTTGTGGCTTTTGAATATCTTCCACAG
AGGGAAGTTTAAAACCCAAACTTCCAAAGGTTT
AAACTACCTCAAAACACTTTCCCATGAGTGTGAT
CCACATTGTTAGGTGCTGACCTAGACAGAGATG
AACTGAGGTCCTTGTTTTGTTTTGTTCATAATAC
AAAGGTGCTAATTAATAGTATTTCAGATACTTGA
AGAATGTTGATGGTGCTAGAAGAATTTGAGAAG
AAATACTCCTGTATTGAGTTGTATCGTGTGGTGT
ATTTTTTAAAAAATTTGATTTAGCATTCATATTTT
CCATCTTATTCCCAATTAAAAGTATGCAGATTAT
TTGCCCAAATCTTCTTCAGATTCAGCATTTGTTCT
TTGCCAGTCTCATTTTCATCTTCTTCCATGGTTCC
ACAGAAGCTTTGTTTCTTGGGCAAGCAGAAAAA
TTAAATTGTACCTATTTTGTATATGTGAGATGTT
TAAATAAATTGTGAAAAAAATGAAATAAAGCAT
GTTTGGTTTTCCAAAAGAACATAT
3UTR-008Col6a2;CGCCGCCGCCCGGGCCCCGCAGTCGAGGGTCGT12
collagen,GAGCCCACCCCGTCCATGGTGCTAAGCGGGCCC
type VI,GGGTCCCACACGGCCAGCACCGCTGCTCACTCG
alpha 2GACGACCCCCTGGGCCTGCACCTCTCCAGCTCCT
CCCACGGGGTCCCCGTAGCCCCGGCCCCCGCCC
AGCCCCAGGTCTCCCCAGGCCCTCCGCAGGCTG
CCCGGCCTCCCTCCCCCTGCAGCCATCCCAAGGC
TCCTGACCTACCTGGCCCCTGAGCTCTGGAGCAA
GCCCTGACCCAATAAAGGCTTTGAACCCAT
3UTR-009RPN1;GGGGCTAGAGCCCTCTCCGCACAGCGTGGAGAC13
ribophorin IGGGGCAAGGAGGGGGGTTATTAGGATTGGTGGT
TTTGTTTTGCTTTGTTTAAAGCCGTGGGAAAATG
GCACAACTTTACCTCTGTGGGAGATGCAACACT
GAGAGCCAAGGGGTGGGAGTTGGGATAATTTTT
ATATAAAAGAAGTTTTTCCACTTTGAATTGCTAA
AAGTGGCATTTTTCCTATGTGCAGTCACTCCTCT
CATTTCTAAAATAGGGACGTGGCCAGGCACGGT
GGCTCATGCCTGTAATCCCAGCACTTTGGGAGGC
CGAGGCAGGCGGCTCACGAGGTCAGGAGATCGA
GACTATCCTGGCTAACACGGTAAAACCCTGTCTC
TACTAAAAGTACAAAAAATTAGCTGGGCGTGGT
GGTGGGCACCTGTAGTCCCAGCTACTCGGGAGG
CTGAGGCAGGAGAAAGGCATGAATCCAAGAGG
CAGAGCTTGCAGTGAGCTGAGATCACGCCATTG
CACTCCAGCCTGGGCAACAGTGTTAAGACTCTGT
CTCAAATATAAATAAATAAATAAATAAATAAAT
AAATAAATAAAAATAAAGCGAGATGTTGCCCTC
AAA
3UTR-010LRP1; lowGGCCCTGCCCCGTCGGACTGCCCCCAGAAAGCC14
densityTCCTGCCCCCTGCCAGTGAAGTCCTTCAGTGAGC
lipoproteinCCCTCCCCAGCCAGCCCTTCCCTGGCCCCGCCGG
receptor-ATGTATAAATGTAAAAATGAAGGAATTACATTT
relatedTATATGTGAGCGAGCAAGCCGGCAAGCGAGCAC
protein 1AGTATTATTTCTCCATCCCCTCCCTGCCTGCTCCT
TGGCACCCCCATGCTGCCTTCAGGGAGACAGGC
AGGGAGGGCTTGGGGCTGCACCTCCTACCCTCC
CACCAGAACGCACCCCACTGGGAGAGCTGGTGG
TGCAGCCTTCCCCTCCCTGTATAAGACACTTTGC
CAAGGCTCTCCCCTCTCGCCCCATCCCTGCTTGC
CCGCTCCCACAGCTTCCTGAGGGCTAATTCTGGG
AAGGGAGAGTTCTTTGCTGCCCCTGTCTGGAAG
ACGTGGCTCTGGGTGAGGTAGGCGGGAAAGGAT
GGAGTGTTTTAGTTCTTGGGGGAGGCCACCCCA
AACCCCAGCCCCAACTCCAGGGGCACCTATGAG
ATGGCCATGCTCAACCCCCCTCCCAGACAGGCC
CTCCCTGTCTCCAGGGCCCCCACCGAGGTTCCCA
GGGCTGGAGACTTCCTCTGGTAAACATTCCTCCA
GCCTCCCCTCCCCTGGGGACGCCAAGGAGGTGG
GCCACACCCAGGAAGGGAAAGCGGGCAGCCCC
GTTTTGGGGACGTGAACGTTTTAATAATTTTTGC
TGAATTCCTTTACAACTAAATAACACAGATATTG
TTATAAATAAAATTGT
3UTR-011Nnt1;ATATTAAGGATCAAGCTGTTAGCTAATAATGCC15
cardiotrophin-ACCTCTGCAGTTTTGGGAACAGGCAAATAAAGT
likeATCAGTATACATGGTGATGTACATCTGTAGCAA
cytokineAGCTCTTGGAGAAAATGAAGACTGAAGAAAGCA
factor 1AAGCAAAAACTGTATAGAGAGATTTTTCAAAAG
CAGTAATCCCTCAATTTTAAAAAAGGATTGAAA
ATTCTAAATGTCTTTCTGTGCATATTTTTTGTGTT
AGGAATCAAAAGTATTTTATAAAAGGAGAAAGA
ACAGCCTCATTTTAGATGTAGTCCTGTTGGATTT
TTTATGCCTCCTCAGTAACCAGAAATGTTTTAAA
AAACTAAGTGTTTAGGATTTCAAGACAACATTAT
ACATGGCTCTGAAATATCTGACACAATGTAAAC
ATTGCAGGCACCTGCATTTTATGTTTTTTTTTTCA
ACAAATGTGACTAATTTGAAACTTTTATGAACTT
CTGAGCTGTCCCCTTGCAATTCAACCGCAGTTTG
AATTAATCATATCAAATCAGTTTTAATTTTTTAA
ATTGTACTTCAGAGTCTATATTTCAAGGGCACAT
TTTCTCACTACTATTTTAATACATTAAAGGACTA
AATAATCTTTCAGAGATGCTGGAAACAAATCAT
TTGCTTTATATGTTTCATTAGAATACCAATGAAA
CATACAACTTGAAAATTAGTAATAGTATTTTTGA
AGATCCCATTTCTAATTGGAGATCTCTTTAATTT
CGATCAACTTATAATGTGTAGTACTATATTAAGT
GCACTTGAGTGGAATTCAACATTTGACTAATAA
AATGAGTTCATCATGTTGGCAAGTGATGTGGCA
ATTATCTCTGGTGACAAAAGAGTAAAATCAAAT
ATTTCTGCCTGTTACAAATATCAAGGAAGACCTG
CTACTATGAAATAGATGACATTAATCTGTCTTCA
CTGTTTATAATACGGATGGATTTTTTTTCAAATC
AGTGTGTGTTTTGAGGTCTTATGTAATTGATGAC
ATTTGAGAGAAATGGTGGCTTTTTTTAGCTACCT
CTTTGTTCATTTAAGCACCAGTAAAGATCATGTC
TTTTTATAGAAGTGTAGATTTTCTTTGTGACTTTG
CTATCGTGCCTAAAGCTCTAAATATAGGTGAATG
TGTGATGAATACTCAGATTATTTGTCTCTCTATA
TAATTAGTTTGGTACTAAGTTTCTCAAAAAATTA
TTAACACATGAAAGACAATCTCTAAACCAGAAA
AAGAAGTAGTACAAATTTTGTTACTGTAATGCTC
GCGTTTAGTGAGTTTAAAACACACAGTATCTTTT
GGTTTTATAATCAGTTTCTATTTTGCTGTGCCTGA
GATTAAGATCTGTGTATGTGTGTGTGTGTGTGTG
TGCGTTTGTGTGTTAAAGCAGAAAAGACTTTTTT
AAAAGTTTTAAGTGATAAATGCAATTTGTTAATT
GATCTTAGATCACTAGTAAACTCAGGGCTGAATT
ATACCATGTATATTCTATTAGAAGAAAGTAAAC
ACCATCTTTATTCCTGCCCTTTTTCTTCTCTCAAA
GTAGTTGTAGTTATATCTAGAAAGAAGCAATTTT
GATTTCTTGAAAAGGTAGTTCCTGCACTCAGTTT
AAACTAAAAATAATCATACTTGGATTTTATTTAT
TTTTGTCATAGTAAAAATTTTAATTTATATATATT
TTTATTTAGTATTATCTTATTCTTTGCTATTTGCC
AATCCTTTGTCATCAATTGTGTTAAATGAATTGA
AAATTCATGCCCTGTTCATTTTATTTTACTTTATT
GGTTAGGATATTTAAAGGATTTTTGTATATATAA
TTTCTTAAATTAATATTCCAAAAGGTTAGTGGAC
TTAGATTATAAATTATGGCAAAAATCTAAAAAC
AACAAAAATGATTTTTATACATTCTATTTCATTA
TTCCTCTTTTTCCAATAAGTCATACAATTGGTAG
ATATGACTTATTTTATTTTTGTATTATTCACTATA
TCTTTATGATATTTAAGTATAAATAATTAAAAAA
ATTTATTGTACCTTATAGTCTGTCACCAAAAAAA
AAAAATTATCTGTAGGTAGTGAAATGCTAATGTT
GATTTGTCTTTAAGGGCTTGTTAACTATCCTTTAT
TTTCTCATTTGTCTTAAATTAGGAGTTTGTGTTTA
AATTACTCATCTAAGCAAAAAATGTATATAAAT
CCCATTACTGGGTATATACCCAAAGGATTATAA
ATCATGCTGCTATAAAGACACATGCACACGTAT
GTTTATTGCAGCACTATTCACAATAGCAAAGACT
TGGAACCAACCCAAATGTCCATCAATGATAGAC
TTGATTAAGAAAATGTGCACATATACACCATGG
AATACTATGCAGCCATAAAAAAGGATGAGTTCA
TGTCCTTTGTAGGGACATGGATAAAGCTGGAAA
CCATCATTCTGAGCAAACTATTGCAAGGACAGA
AAACCAAACACTGCATGTTCTCACTCATAGGTG
GGAATTGAACAATGAGAACACTTGGACACAAGG
TGGGGAACACCACACACCAGGGCCTGTCATGGG
GTGGGGGGAGTGGGGAGGGATAGCATTAGGAG
ATATACCTAATGTAAATGATGAGTTAATGGGTG
CAGCACACCAACATGGCACATGTATACATATGT
AGCAAACCTGCACGTTGTGCACATGTACCCTAG
AACTTAAAGTATAATTAAAAAAAAAAAGAAAAC
AGAAGCTATTTATAAAGAAGTTATTTGCTGAAAT
AAATGTGATCTTTCCCATTAAAAAAATAAAGAA
ATTTTGGGGTAAAAAAACACAATATATTGTATTC
TTGAAAAATTCTAAGAGAGTGGATGTGAAGTGT
TCTCACCACAAAAGTGATAACTAATTGAGGTAA
TGCACATATTAATTAGAAAGATTTTGTCATTCCA
CAATGTATATATACTTAAAAATATGTTATACACA
ATAAATACATACATTAAAAAATAAGTAAATGTA
3UTR-012Col6a1;CCCACCCTGCACGCCGGCACCAAACCCTGTCCTC16
collagen,CCACCCCTCCCCACTCATCACTAAACAGAGTAA
type VI,AATGTGATGCGAATTTTCCCGACCAACCTGATTC
alpha 1GCTAGATTTTTTTTAAGGAAAAGCTTGGAAAGCC
AGGACACAACGCTGCTGCCTGCTTTGTGCAGGG
TCCTCCGGGGCTCAGCCCTGAGTTGGCATCACCT
GCGCAGGGCCCTCTGGGGCTCAGCCCTGAGCTA
GTGTCACCTGCACAGGGCCCTCTGAGGCTCAGC
CCTGAGCTGGCGTCACCTGTGCAGGGCCCTCTGG
GGCTCAGCCCTGAGCTGGCCTCACCTGGGTTCCC
CACCCCGGGCTCTCCTGCCCTGCCCTCCTGCCCG
CCCTCCCTCCTGCCTGCGCAGCTCCTTCCCTAGG
CACCTCTGTGCTGCATCCCACCAGCCTGAGCAAG
ACGCCCTCTCGGGGCCTGTGCCGCACTAGCCTCC
CTCTCCTCTGTCCCCATAGCTGGTTTTTCCCACCA
ATCCTCACCTAACAGTTACTTTACAATTAAACTC
AAAGCAAGCTCTTCTCCTCAGCTTGGGGCAGCC
ATTGGCCTCTGTCTCGTTTTGGGAAACCAAGGTC
AGGAGGCCGTTGCAGACATAAATCTCGGCGACT
CGGCCCCGTCTCCTGAGGGTCCTGCTGGTGACCG
GCCTGGACCTTGGCCCTACAGCCCTGGAGGCCG
CTGCTGACCAGCACTGACCCCGACCTCAGAGAG
TACTCGCAGGGGCGCTGGCTGCACTCAAGACCC
TCGAGATTAACGGTGCTAACCCCGTCTGCTCCTC
CCTCCCGCAGAGACTGGGGCCTGGACTGGACAT
GAGAGCCCCTTGGTGCCACAGAGGGCTGTGTCT
TACTAGAAACAACGCAAACCTCTCCTTCCTCAGA
ATAGTGATGTGTTCGACGTTTTATCAAAGGCCCC
CTTTCTATGTTCATGTTAGTTTTGCTCCTTCTGTG
TTTTTTTCTGAACCATATCCATGTTGCTGACTTTT
CCAAATAAAGGTTTTCACTCCTCTC
3UTR-013Calr;AGAGGCCTGCCTCCAGGGCTGGACTGAGGCCTG17
calreticulinAGCGCTCCTGCCGCAGAGCTGGCCGCGCCAAAT
AATGTCTCTGTGAGACTCGAGAACTTTCATTTTT
TTCCAGGCTGGTTCGGATTTGGGGTGGATTTTGG
TTTTGTTCCCCTCCTCCACTCTCCCCCACCCCCTC
CCCGCCCTTTTTTTTTTTTTTTTTTAAACTGGTAT
TTTATCTTTGATTCTCCTTCAGCCCTCACCCCTGG
TTCTCATCTTTCTTGATCAACATCTTTTCTTGCCT
CTGTCCCCTTCTCTCATCTCTTAGCTCCCCTCCAA
CCTGGGGGGCAGTGGTGTGGAGAAGCCACAGGC
CTGAGATTTCATCTGCTCTCCTTCCTGGAGCCCA
GAGGAGGGCAGCAGAAGGGGGTGGTGTCTCCAA
CCCCCCAGCACTGAGGAAGAACGGGGCTCTTCT
CATTTCACCCCTCCCTTTCTCCCCTGCCCCCAGG
ACTGGGCCACTTCTGGGTGGGGCAGTGGGTCCC
AGATTGGCTCACACTGAGAATGTAAGAACTACA
AACAAAATTTCTATTAAATTAAATTTTGTGTCTCC
3UTR-014Colla1;CTCCCTCCATCCCAACCTGGCTCCCTCCCACCCA18
collagen,ACCAACTTTCCCCCCAACCCGGAAACAGACAAG
type I,CAACCCAAACTGAACCCCCTCAAAAGCCAAAAA
alpha 1ATGGGAGACAATTTCACATGGACTTTGGAAAAT
ATTTTTTTCCTTTGCATTCATCTCTCAAACTTAGT
TTTTATCTTTGACCAACCGAACATGACCAAAAAC
CAAAAGTGCATTCAACCTTACCAAAAAAAAAAA
AAAAAAAAGAATAAATAAATAACTTTTTAAAAA
AGGAAGCTTGGTCCACTTGCTTGAAGACCCATG
CGGGGGTAAGTCCCTTTCTGCCCGTTGGGCTTAT
GAAACCCCAATGCTGCCCTTTCTGCTCCTTTCTC
CACACCCCCCTTGGGGCCTCCCCTCCACTCCTTC
CCAAATCTGTCTCCCCAGAAGACACAGGAAACA
ATGTATTGTCTGCCCAGCAATCAAAGGCAATGCT
CAAACACCCAAGTGGCCCCCACCCTCAGCCCGC
TCCTGCCCGCCCAGCACCCCCAGGCCCTGGGGG
ACCTGGGGTTCTCAGACTGCCAAAGAAGCCTTG
CCATCTGGCGCTCCCATGGCTCTTGCAACATCTC
CCCTTCGTTTTTGAGGGGGTCATGCCGGGGGAGC
CACCAGCCCCTCACTGGGTTCGGAGGAGAGTCA
GGAAGGGCCACGACAAAGCAGAAACATCGGATT
TGGGGAACGCGTGTCAATCCCTTGTGCCGCAGG
GCTGGGCGGGAGAGACTGTTCTGTTCCTTGTGTA
ACTGTGTTGCTGAAAGACTACCTCGTTCTTGTCT
TGATGTGTCACCGGGGCAACTGCCTGGGGGCGG
GGATGGGGGCAGGGTGGAAGCGGCTCCCCATTT
TATACCAAAGGTGCTACATCTATGTGATGGGTG
GGGTGGGGAGGGAATCACTGGTGCTATAGAAAT
TGAGATGCCCCCCCAGGCCAGCAAATGTTCCTTT
TTGTTCAAAGTCTATTTTTATTCCTTGATATTTTT
CTTTTTTTTTTTTTTTTTTTGTGGATGGGGACTTG
TGAATTTTTCTAAAGGTGCTATTTAACATGGGAG
GAGAGCGTGTGCGGCTCCAGCCCAGCCCGCTGC
TCACTTTCCACCCTCTCTCCACCTGCCTCTGGCTT
CTCAGGCCTCTGCTCTCCGACCTCTCTCCTCTGA
AACCCTCCTCCACAGCTGCAGCCCATCCTCCCGG
CTCCCTCCTAGTCTGTCCTGCGTCCTCTGTCCCCG
GGTTTCAGAGACAACTTCCCAAAGCACAAAGCA
GTTTTTCCCCCTAGGGGTGGGAGGAAGCAAAAG
ACTCTGTACCTATTTTGTATGTGTATAATAATTT
GAGATGTTTTTAATTATTTTGATTGCTGGAATAA
AGCATGTGGAAATGACCCAAACATAATCCGCAG
TGGCCTCCTAATTTCCTTCTTTGGAGTTGGGGGA
GGGGTAGACATGGGGAAGGGGCTTTGGGGTGAT
GGGCTTGCCTTCCATTCCTGCCCTTTCCCTCCCCA
CTATTCTCTTCTAGATCCCTCCATAACCCCACTC
CCCTTTCTCTCACCCTTCTTATACCGCAAACCTTT
CTACTTCCTCTTTCATTTTCTATTCTTGCAATTTC
CTTGCACCTTTTCCAAATCCTCTTCTCCCCTGCAA
TACCATACAGGCAATCCACGTGCACAACACACA
CACACACTCTTCACATCTGGGGTTGTCCAAACCT
CATACCCACTCCCCTTCAAGCCCATCCACTCTCC
ACCCCCTGGATGCCCTGCACTTGGTGGCGGTGG
GATGCTCATGGATACTGGGAGGGTGAGGGGAGT
GGAACCCGTGAGGAGGACCTGGGGGCCTCTCCT
TGAACTGACATGAAGGGTCATCTGGCCTCTGCTC
CCTTCTCACCCACGCTGACCTCCTGCCGAAGGAG
CAACGCAACAGGAGAGGGGTCTGCTGAGCCTGG
CGAGGGTCTGGGAGGGACCAGGAGGAAGGCGT
GCTCCCTGCTCGCTGTCCTGGCCCTGGGGGAGTG
AGGGAGACAGACACCTGGGAGAGCTGTGGGGA
AGGCACTCGCACCGTGCTCTTGGGAAGGAAGGA
GACCTGGCCCTGCTCACCACGGACTGGGTGCCTC
GACCTCCTGAATCCCCAGAACACAACCCCCCTG
GGCTGGGGTGGTCTGGGGAACCATCGTGCCCCC
GCCTCCCGCCTACTCCTTTTTAAGCTT
3UTR-015Plod1;TTGGCCAGGCCTGACCCTCTTGGACCTTTCTTCT19
procollagen-TTGCCGACAACCACTGCCCAGCAGCCTCTGGGA
lysine, 2-CCTCGGGGTCCCAGGGAACCCAGTCCAGCCTCC
oxoglutarateTGGCTGTTGACTTCCCATTGCTCTTGGAGCCACC
5-AATCAAAGAGATTCAAAGAGATTCCTGCAGGCC
dioxygenase 1AGAGGCGGAACACACCTTTATGGCTGGGGCTCT
CCGTGGTGTTCTGGACCCAGCCCCTGGAGACAC
CATTCACTTTTACTGCTTTGTAGTGACTCGTGCTC
TCCAACCTGTCTTCCTGAAAAACCAAGGCCCCCT
TCCCCCACCTCTTCCATGGGGTGAGACTTGAGCA
GAACAGGGGCTTCCCCAAGTTGCCCAGAAAGAC
TGTCTGGGTGAGAAGCCATGGCCAGAGCTTCTC
CCAGGCACAGGTGTTGCACCAGGGACTTCTGCTT
CAAGTTTTGGGGTAAAGACACCTGGATCAGACT
CCAAGGGCTGCCCTGAGTCTGGGACTTCTGCCTC
CATGGCTGGTCATGAGAGCAAACCGTAGTCCCC
TGGAGACAGCGACTCCAGAGAACCTCTTGGGAG
ACAGAAGAGGCATCTGTGCACAGCTCGATCTTC
TACTTGCCTGTGGGGAGGGGAGTGACAGGTCCA
CACACCACACTGGGTCACCCTGTCCTGGATGCCT
CTGAAGAGAGGGACAGACCGTCAGAAACTGGA
GAGTTTCTATTAAAGGTCATTTAAACCA
3UTR-016Nucb1;TCCTCCGGGACCCCAGCCCTCAGGATTCCTGATG20
nucleobindin 1CTCCAAGGCGACTGATGGGCGCTGGATGAAGTG
GCACAGTCAGCTTCCCTGGGGGCTGGTGTCATGT
TGGGCTCCTGGGGCGGGGGCACGGCCTGGCATT
TCACGCATTGCTGCCACCCCAGGTCCACCTGTCT
CCACTTTCACAGCCTCCAAGTCTGTGGCTCTTCC
CTTCTGTCCTCCGAGGGGCTTGCCTTCTCTCGTG
TCCAGTGAGGTGCTCAGTGATCGGCTTAACTTAG
AGAAGCCCGCCCCCTCCCCTTCTCCGTCTGTCCC
AAGAGGGTCTGCTCTGAGCCTGCGTTCCTAGGTG
GCTCGGCCTCAGCTGCCTGGGTTGTGGCCGCCCT
AGCATCCTGTATGCCCACAGCTACTGGAATCCCC
GCTGCTGCTCCGGGCCAAGCTTCTGGTTGATTAA
TGAGGGCATGGGGTGGTCCCTCAAGACCTTCCC
CTACCTTTTGTGGAACCAGTGATGCCTCAAAGAC
AGTGTCCCCTCCACAGCTGGGTGCCAGGGGCAG
GGGATCCTCAGTATAGCCGGTGAACCCTGATAC
CAGGAGCCTGGGCCTCCCTGAACCCCTGGCTTCC
AGCCATCTCATCGCCAGCCTCCTCCTGGACCTCT
TGGCCCCCAGCCCCTTCCCCACACAGCCCCAGA
AGGGTCCCAGAGCTGACCCCACTCCAGGACCTA
GGCCCAGCCCCTCAGCCTCATCTGGAGCCCCTGA
AGACCAGTCCCACCCACCTTTCTGGCCTCATCTG
ACACTGCTCCGCATCCTGCTGTGTGTCCTGTTCC
ATGTTCCGGTTCCATCCAAATACACTTTCTGGAA
CAAA
3UTR-017α-globinGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTT21
GGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCA
CCCGTACCCCCGTGGTCTTTGAATAAAGTCTGAG
TGGGCGGC

It should be understood that those listed in the previous tables are examples and that any UTR from any gene may be incorporated into the respective first or second flanking region of the primary construct. Furthermore, multiple wild-type UTRs of any known gene may be utilized. It is also within the scope of the present invention to provide artificial UTRs which are not variants of wild type genes. These UTRs or portions thereof may be placed in the same orientation as in the transcript from which they were selected or may be altered in orientation or location. Hence a 5′ or 3′ UTR may be inverted, shortened, lengthened, made chimeric with one or more other 5′ UTRs or 3′ UTRs. As used herein, the term “altered” as it relates to a UTR sequence, means that the UTR has been changed in some way in relation to a reference sequence. For example, a 3′ or 5′ UTR may be altered relative to a wild type or native UTR by the change in orientation or location as taught above or may be altered by the inclusion of additional nucleotides, deletion of nucleotides, swapping or transposition of nucleotides. Any of these changes producing an “altered” UTR (whether 3′ or 5′) comprise a variant UTR.

In one embodiment, a double, triple or quadruple UTR such as a 5′ or 3′ UTR may be used. As used herein, a “double” UTR is one in which two copies of the same UTR are encoded either in series or substantially in series. For example, a double beta-globin 3′ UTR may be used as described in US Patent publication 20100129877, the contents of which are incorporated herein by reference in its entirety.

It is also within the scope of the present invention to have patterned UTRs. As used herein “patterned UTRs” are those UTRs which reflect a repeating or alternating pattern, such as ABABAB or AABBAABBAABB or ABCABCABC or variants thereof repeated once, twice, or more than 3 times. In these patterns, each letter, A, B, or C represent a different UTR at the nucleotide level.

In one embodiment, flanking regions are selected from a family of transcripts whose proteins share a common function, structure, feature of property. For example, oncology-related polypeptides of interest may belong to a family of proteins which are expressed in a particular cell, tissue or at some time during development. The UTRs from any of these genes may be swapped for any other UTR of the same or different family of proteins to create a new chimeric primary transcript. As used herein, a “family of proteins” is used in the broadest sense to refer to a group of two or more oncology-related polypeptides of interest which share at least one function, structure, feature, localization, origin, or expression pattern.

After optimization (if desired), the signal-sensor primary construct components are reconstituted and transformed into a vector such as, but not limited to, plasmids, viruses, cosmids, and artificial chromosomes. For example, the optimized construct may be reconstituted and transformed into chemically competent E. coli, yeast, neurospora, maize, drosophila, etc. where high copy plasmid-like or chromosome structures occur by methods described herein.

Stop Codons

In one embodiment, the signal-sensor primary constructs of the present invention may include at least two stop codons before the 3′ untranslated region (UTR). The stop codon may be selected from TGA, TAA and TAG. In one embodiment, the signal-sensor primary constructs of the present invention include the stop codon TGA and one additional stop codon. In a further embodiment the addition stop codon may be TAA.

Vector Amplification

The vector containing the signal-sensor primary construct is then amplified and the plasmid isolated and purified using methods known in the art such as, but not limited to, a maxi prep using the Invitrogen PURELINK™ HiPure Maxiprep Kit (Carlsbad, Calif.).

Plasmid Linearization

The plasmid may then be linearized using methods known in the art such as, but not limited to, the use of restriction enzymes and buffers. The linearization reaction may be purified using methods including, for example Invitrogen's PURELINK™ PCR Micro Kit (Carlsbad, Calif.), and HPLC based purification methods such as, but not limited to, strong anion exchange HPLC, weak anion exchange HPLC, reverse phase HPLC (RP-HPLC), and hydrophobic interaction HPLC (HIC-HPLC) and Invitrogen's standard PURELINK™ PCR Kit (Carlsbad, Calif.). The purification method may be modified depending on the size of the linearization reaction which was conducted. The linearized plasmid is then used to generate cDNA for in vitro transcription (IVT) reactions.

cDNA Template Synthesis

A cDNA template may be synthesized by having a linearized plasmid undergo polymerase chain reaction (PCR). Table 4 is a listing of primers and probes that may be useful in the PCR reactions of the present invention. It should be understood that the listing is not exhaustive and that primer-probe design for any amplification is within the skill of those in the art. Probes may also contain chemically modified bases to increase base-pairing fidelity to the target molecule and base-pairing strength. Such modifications may include 5-methyl-Cytidine, 2,6-di-amino-purine, 2′-fluoro, phosphoro-thioate, or locked nucleic acids.

TABLE 4
Primers and Probes
Primer/SEQ
ProbeHybridizationID
IdentifierSequence (5′-3′)targetNO.
UFPTTGGACCCTCGTACAGAAGCTAAcDNA Template22
TACG
URPTx160CTTCCTACTCAGGCTTTATTCcDNA Template23
AAAGACCA
GBA1CCTTGACCTTCTGGAACTTCAcid24
glucocerebrosidase
GBA2CCAAGCACTGAAACGGATATAcid25
glucocerebrosidase
LUC1GATGAAAAGTGCTCCAAGGALuciferase26
LUC2AACCGTGATGAAAAGGTACCLuciferase27
LUC3TCATGCAGATTGGAAAGGTCLuciferase28
GCSF1CTTCTTGGACTGTCCAGAGGG-CSF29
GCSF2GCAGTCCCTGATACAAGAACG-CSF30
GCSF3GATTGAAGGTGGCTCGCTACG-CSF31
*UFP is universal forward primer; URP is universal reverse primer.

In one embodiment, the cDNA may be submitted for sequencing analysis before undergoing transcription.

Signal-Sensor Polynucleotide Production (Signal-Sensor mRNA)

The process of signal-sensor polynucleotide production may include, but is not limited to, in vitro transcription, cDNA template removal and RNA clean-up, and capping and/or tailing reactions.

In Vitro Transcription

The cDNA produced in the previous step may be transcribed using an in vitro transcription (IVT) system. The system typically comprises a transcription buffer, nucleotide triphosphates (NTPs), an RNase inhibitor and a polymerase. The NTPs may be manufactured in house, may be selected from a supplier, or may be synthesized as described herein. The NTPs may be selected from, but are not limited to, those described herein including natural and unnatural (modified) NTPs. The polymerase may be selected from, but is not limited to, T7 RNA polymerase, T3 RNA polymerase and mutant polymerases such as, but not limited to, polymerases able to be incorporated into modified nucleic acids.

RNA Polymerases

Any number of RNA polymerases or variants may be used in the design of the signal-sensor primary constructs of the present invention.

RNA polymerases may be modified by inserting or deleting amino acids of the RNA polymerase sequence. As a non-limiting example, the RNA polymerase may be modified to exhibit an increased ability to incorporate a 2′-modified nucleotide triphosphate compared to an unmodified RNA polymerase (see International Publication WO2008078180 and U.S. Pat. No. 8,101,385; herein incorporated by reference in their entireties).

Variants may be obtained by evolving an RNA polymerase, optimizing the RNA polymerase amino acid and/or nucleic acid sequence and/or by using other methods known in the art. As a non-limiting example, T7 RNA polymerase variants may be evolved using the continuous directed evolution system set out by Esvelt et al. (Nature (2011) 472(7344):499-503; herein incorporated by reference in its entirety) where clones of T7 RNA polymerase may encode at least one mutation such as, but not limited to, lysine at position 93 substituted for threonine (K93T), I4M, A7T, E63V, V64D, A65E, D66Y, T76N, C125R, S128R, A136T, N165S, G175R, H176L, Y178H, F182L, L196F, G198V, D208Y, E222K, S228A, Q239R, T243N, G259D, M267I, G280C, H300R, D351A, A354S, E356D, L360P, A383V, Y385C, D388Y, S397R, M401T, N410S, K450R, P451T, G452V, E484A, H523L, H524N, G542V, E565K, K577E, K577M, N601S, S684Y, L699I, K713E, N748D, Q754R, E775K, A827V, D851N or L864F. As another non-limiting example, T7 RNA polymerase variants may encode at least mutation as described in U.S. Pub. Nos. 20100120024 and 20070117112; herein incorporated by reference in their entireties. Variants of RNA polymerase may also include, but are not limited to, substitutional variants, conservative amino acid substitution, insertional variants, deletional variants and/or covalent derivatives.

In one embodiment, the signal-sensor primary construct may be designed to be recognized by the wild type or variant RNA polymerases. In doing so, the signal-sensor primary construct may be modified to contain sites or regions of sequence changes from the wild type or parent primary construct.

In one embodiment, the signal-sensor primary construct may be designed to include at least one substitution and/or insertion upstream of an RNA polymerase binding or recognition site, downstream of the RNA polymerase binding or recognition site, upstream of the TATA box sequence, downstream of the TATA box sequence of the signal-sensor primary construct but upstream of the coding region of the primary construct, within the 5′UTR, before the 5′UTR and/or after the 5′UTR.

In one embodiment, the 5′UTR of the signal-sensor primary construct may be replaced by the insertion of at least one region and/or string of nucleotides of the same base. The region and/or string of nucleotides may include, but is not limited to, at least 3, at least 4, at least 5, at least 6, at least 7 or at least 8 nucleotides and the nucleotides may be natural and/or unnatural. As a non-limiting example, the group of nucleotides may include 5-8 adenine, cytosine, thymine, a string of any of the other nucleotides disclosed herein and/or combinations thereof.

In one embodiment, the 5′UTR of the signal-sensor primary construct may be replaced by the insertion of at least two regions and/or strings of nucleotides of two different bases such as, but not limited to, adenine, cytosine, thymine, any of the other nucleotides disclosed herein and/or combinations thereof. For example, the 5′UTR may be replaced by inserting 5-8 adenine bases followed by the insertion of 5-8 cytosine bases. In another example, the 5′UTR may be replaced by inserting 5-8 cytosine bases followed by the insertion of 5-8 adenine bases.

In one embodiment, the signal-sensor primary construct may include at least one substitution and/or insertion downstream of the transcription start site which may be recognized by an RNA polymerase. As a non-limiting example, at least one substitution and/or insertion may occur downstream the transcription start site by substituting at least one nucleic acid in the region just downstream of the transcription start site (such as, but not limited to, +1 to +6). Changes to region of nucleotides just downstream of the transcription start site may affect initiation rates, increase apparent nucleotide triphosphate (NTP) reaction constant values, and increase the dissociation of short transcripts from the transcription complex curing initial transcription (Brieba et al, Biochemistry (2002) 41: 5144-5149; herein incorporated by reference in its entirety). The modification, substitution and/or insertion of at least one nucleic acid may cause a silent mutation of the nucleic acid sequence or may cause a mutation in the amino acid sequence.

In one embodiment, the signal-sensor primary construct may include the substitution of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12 or at least 13 guanine bases downstream of the transcription start site.

In one embodiment, the signal-sensor primary construct may include the substitution of at least 1, at least 2, at least 3, at least 4, at least 5 or at least 6 guanine bases in the region just downstream of the transcription start site. As a non-limiting example, if the nucleotides in the region are GGGAGA the guanine bases may be substituted by at least 1, at least 2, at least 3 or at least 4 adenine nucleotides. In another non-limiting example, if the nucleotides in the region are GGGAGA the guanine bases may be substituted by at least 1, at least 2, at least 3 or at least 4 cytosine bases. In another non-limiting example, if the nucleotides in the region are GGGAGA the guanine bases may be substituted by at least 1, at least 2, at least 3 or at least 4 thymine, and/or any of the nucleotides described herein.

In one embodiment, the signal-sensor primary construct may include at least one substitution and/or insertion upstream of the start codon. For the purpose of clarity, one of skill in the art would appreciate that the start codon is the first codon of the protein coding region whereas the transcription start site is the site where transcription begins. The signal-sensor primary construct may include, but is not limited to, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7 or at least 8 substitutions and/or insertions of nucleotide bases. The nucleotide bases may be inserted or substituted at 1, at least 1, at least 2, at least 3, at least 4 or at least 5 locations upstream of the start codon. The nucleotides inserted and/or substituted may be the same base (e.g., all A or all C or all T or all G), two different bases (e.g., A and C, A and T, or C and T), three different bases (e.g., A, C and T or A, C and T) or at least four different bases. As a non-limiting example, the guanine base upstream of the coding region in the signal-sensor primary construct may be substituted with adenine, cytosine, thymine, or any of the nucleotides described herein. In another non-limiting example the substitution of guanine bases in the signal-sensor primary construct may be designed so as to leave one guanine base in the region downstream of the transcription start site and before the start codon (see Esvelt et al. Nature (2011) 472(7344):499-503; herein incorporated by reference in its entirety). As a non-limiting example, at least 5 nucleotides may be inserted at 1 location downstream of the transcription start site but upstream of the start codon and the at least 5 nucleotides may be the same base type.

cDNA Template Removal and Clean-Up

The cDNA template may be removed using methods known in the art such as, but not limited to, treatment with Deoxyribonuclease I (DNase I). RNA clean-up may also include a purification method such as, but not limited to, AGENCOURT® CLEANSEQ® system from Beckman Coulter (Danvers, Mass.), HPLC based purification methods such as, but not limited to, strong anion exchange HPLC, weak anion exchange HPLC, reverse phase HPLC (RP-HPLC), and hydrophobic interaction HPLC (HIC-HPLC).

Capping and/or Tailing Reactions

The signal-sensor primary construct or mmRNA may also undergo capping and/or tailing reactions. A capping reaction may be performed by methods known in the art to add a 5′ cap to the 5′ end of the signal-sensor primary construct. Methods for capping include, but are not limited to, using a Vaccinia Capping enzyme (New England Biolabs, Ipswich, Mass.).

A poly-A tailing reaction may be performed by methods known in the art, such as, but not limited to, 2′ O-methyltransferase and by methods as described herein. If the signal-sensor primary construct generated from cDNA does not include a poly-T, it may be beneficial to perform the poly-A-tailing reaction before the signal-sensor primary construct is cleaned.

Purification

Signal-sensor primary construct or mmRNA purification may include, but is not limited to, mRNA or mmRNA clean-up, quality assurance and quality control. mRNA or mmRNA clean-up may be performed by methods known in the arts such as, but not limited to, AGENCOURT® beads (Beckman Coulter Genomics, Danvers, Mass.), poly-T beads, LNA™ oligo-T capture probes (EXIQON® Inc, Vedbaek, Denmark) or HPLC based purification methods such as, but not limited to, strong anion exchange HPLC, weak anion exchange HPLC, reverse phase HPLC (RP-HPLC), and hydrophobic interaction HPLC (HIC-HPLC). The term “purified” when used in relation to a polynucleotide such as a “purified mRNA or signal-sensor mmRNA” refers to one that is separated from at least one contaminant. As used herein, a “contaminant” is any substance which makes another unfit, impure or inferior. Thus, a purified signal-sensor polynucleotide (e.g., DNA and RNA) is present in a form or setting different from that in which it is found in nature, or a form or setting different from that which existed prior to subjecting it to a treatment or purification method.

A quality assurance and/or quality control check may be conducted using methods such as, but not limited to, gel electrophoresis, UV absorbance, or analytical HPLC.

In another embodiment, the signal-sensor mRNA or mmRNA may be sequenced by methods including, but not limited to reverse-transcriptase-PCR.

In one embodiment, the signal-sensor mRNA or mmRNA may be quantified using methods such as, but not limited to, ultraviolet visible spectroscopy (UV/Vis). A non-limiting example of a UV/Vis spectrometer is a NANODROP® spectrometer (ThermoFisher, Waltham, Mass.). The quantified signal-sensor mRNA or mmRNA may be analyzed in order to determine if the signal-sensor mRNA or mmRNA may be of proper size, check that no degradation of the signal-sensor mRNA or mmRNA has occurred. Degradation of the signal-sensor mRNA and/or mmRNA may be checked by methods such as, but not limited to, agarose gel electrophoresis, HPLC based purification methods such as, but not limited to, strong anion exchange HPLC, weak anion exchange HPLC, reverse phase HPLC (RP-HPLC), and hydrophobic interaction HPLC (HIC-HPLC), liquid chromatography-mass spectrometry (LCMS), capillary electrophoresis (CE) and capillary gel electrophoresis (CGE).

Signal Peptides or Proteins

The signal-sensor primary constructs or mmRNA may also encode additional features which facilitate trafficking of the polypeptides to therapeutically relevant sites. One such feature which aids in protein trafficking is the signal peptide sequence. As used herein, a “signal sequence” or “signal peptide” is a polynucleotide or polypeptide, respectively, which is from about 9 to 200 nucleotides (3-60 amino acids) in length which is incorporated at the 5′ (or N-terminus) of the coding region or polypeptide encoded, respectively. Addition of these sequences result in trafficking of the encoded oncology-related polypeptide to the endoplasmic reticulum through one or more secretory pathways. Some signal peptides are cleaved from the protein by signal peptidase after the proteins are transported.

Table 5 is a representative listing of signal proteins or peptides which may be incorporated for encoding by the signal-sensor polynucleotides, primary constructs or mmRNA of the invention.

TABLE 5
Signal Peptides
SEQSEQ
NUCLEOTIDE SEQUENCEIDENCODEDID
IDDescription(5′-3′)NO.PEPTIDENO.
SS-001α-1-ATGATGCCATCCTCAGTCTCA32MMPSSVS94
antitrypsinTGGGGTATTTTGCTCTTGGCGWGILLAGL
GGTCTGTGCTGTCTCGTGCCGCCLVPVSLA
GTGTCGCTCGCA
SS-002G-CSFATGGCCGGACCGGCGACTCAG33MAGPATQ95
TCGCCCATGAAACTCATGGCCSPMKLMA
CTGCAGTTGTTGCTTTGGCACLQLLLWH
TCAGCCCTCTGGACCGTCCAASALWTVQ
GAGGCGEA
SS-003Factor IXATGCAGAGAGTGAACATGATT34MQRVNMI96
ATGGCCGAGTCCCCATCGCTCMAESPSLI
ATCACAATCTGCCTGCTTGGTTICLLGYL
ACCTGCTTTCCGCCGAATGCALSAECTVF
CTGTCTTTCTGGATCACGAGALDHENAN
ATGCGAATAAGATCTTGAACCKILNRPKR
GACCCAAACGG
SS-004ProlactinATGAAAGGATCATTGCTGTTG35MKGSLLL97
CTCCTCGTGTCGAACCTTCTGLLVSNLLL
CTTTGCCAGTCCGTAGCCCCCCQSVAP
SS-005AlbuminATGAAATGGGTGACGTTCATC36MKWVTFI98
TCACTGTTGTTTTTGTTCTCGTSLLFLFSS
CCGCCTACTCCAGGGGAGTATAYSRG
TCCGCCGAVFRR
SS-006HMMSP38ATGTGGTGGCGGCTCTGGTGG37MWWRLW99
CTGCTCCTGTTGCTCCTCTTGCWLLLLLLL
TGTGGCCCATGGTGTGGGCALPMWA
MLS-ornithineTGCTCTTTAACCTCCGCATCCT38MLFNLRIL100
001carbamoyltransferaseGTTGAATAACGCTGCGTTCCGLNNAAFR
AAATGGGCATAACTTCATGGTNGHNFMV
ACGCAACTTCAGATGCGGCCARNFRCGQP
GCCACTCCAGLQ
MLS-CytochromeATGTCCGTCTTGACACCCCTG39MSVLTPLL101
002C OxidaseCTCTTGAGAGGGCTGACGGGGLRGLTGSA
subunit 8ATCCGCTAGACGCCTGCCGGTARRLPVPRA
CCGCGAGCGAAGATCCACTCCKIHSL
CTG
MLS-CytochromeATGAGCGTGCTCACTCCGTTG40MSVLTPLL102
003C OxidaseCTTCTTCGAGGGCTTACGGGALRGLTGSA
subunit 8ATCGGCTCGGAGGTTGCCCGTCRRLPVPRA
CCGAGAGCGAAGATCCATTCGKIHSL
TTG
SS-007Type III,TGACAAAAATAACTTTATCTC41MVTKITLS103
bacterialCCCAGAATTTTAGAATCCAAAPQNFRIQK
AACAGGAAACCACACTACTAQETTLLKE
AAAGAAAAATCAACCGAGAAKSTEKNSL
AAATTCTTTAGCAAAAAGTATAKSILAVK
TCTCGCAGTAAAAATCACTTCNHFIELRS
ATCGAATTAAGGTCAAAATTAKLSERFIS
TCGGAACGTTTTATTTCGCATHKNT
AAGAACACT
SS-008ViralATGCTGAGCTTTGTGGATACC42MLSFVDT104
CGCACCCTGCTGCTGCTGGCGRTLLLLAV
GTGACCAGCTGCCTGGCGACCTSCLATCQ
TGCCAG
SS-009viralATGGGCAGCAGCCAGGCGCC43MGSSQAP105
GCGCATGGGCAGCGTGGGCGRMGSVGG
GCCATGGCCTGATGGCGCTGCHGLMALL
TGATGGCGGGCCTGATTCTGCMAGLILPG
CGGGCATTCTGGCGILA
SS-010ViralATGGCGGGCATTTTTTATTTTC44MAGIFYFL106
TGTTTAGCTTTCTGTTTGGCATFSFLFGICD
TTGCGAT
SS-011ViralATGGAAAACCGCCTGCTGCGC45MENRLLR107
GTGTTTCTGGTGTGGGCGGCGVFLVWAA
CTGACCATGGATGGCGCGAGCLTMDGASA
GCG
SS-012ViralATGGCGCGCCAGGGCTGCTTT46MARQGCF108
GGCAGCTATCAGGTGATTAGCGSYQVISL
CTGTTTACCTTTGCGATTGGCFTFAIGVN
GTGAACCTGTGCCTGGGCLCLG
SS-013BacillusATGAGCCGCCTGCCGGTGCTG47MSRLPVLL109
CTGCTGCTGCAGCTGCTGGTGLLQLLVRP
CGCCCGGGCCTGCAGGLQ
SS-014BacillusATGAAACAGCAGAAACGCCT48MKQQKRL110
GTATGCGCGCCTGCTGACCCTYARLLTLL
GCTGTTTGCGCTGATTTTTCTGFALIFLLPH
CTGCCGCATAGCAGCGCGAGCSSASA
GCG
SS-015SecretionATGGCGACGCCGCTGCCTCCG49MATPLPPP111
signalCCCTCCCCGCGGCACCTGCGGSPRHLRLL
CTGCTGCGGCTGCTGCTCTCCRLLLSG
GCCCTCGTCCTCGGC
SS-016SecretionATGAAGGCTCCGGGTCGGCTC50MKAPGRL112
signalGTGCTCATCATCCTGTGCTCCVLIILCSVV
GTGGTCTTCTCTFS
SS-017SecretionATGCTTCAGCTTTGGAAACTT51MLQLWKL113
signalGTTCTCCTGTGCGGCGTGCTCLCGVLT
ACT
SS-018SecretionATGCTTTATCTCCAGGGTTGG52MLYLQGW114
signalAGCATGCCTGCTGTGGCASMPAVA
SS-019SecretionATGGATAACGTGCAGCCGAA53MDNVQPK115
signalAATAAAACATCGCCCCTTCTGIKHRPFCF
CTTCAGTGTGAAAGGCCACGTSVKGHVK
GAAGATGCTGCGGCTGGATATMLRLDIIN
TATCAACTCACTGGTAACAACSLVTTVFM
AGTATTCATGCTCATCGTATCLIVSVLALIP
TGTGTTGGCACTGATACCA
SS-020SecretionATGCCCTGCCTAGACCAACAG54MPCLDQQ116
signalCTCACTGTTCATGCCCTACCCTLTVHALPC
GCCCTGCCCAGCCCTCCTCTCPAQPSSLA
TGGCCTTCTGCCAAGTGGGGTFCQVGFLTA
TCTTAACAGCA
SS-021SecretionATGAAAACCTTGTTCAATCCA55MKTLFNP117
signalGCCCCTGCCATTGCTGACCTGAPAIADLD
GATCCCCAGTTCTACACCCTCPQFYTLSD
TCAGATGTGTTCTGCTGCAATVFCCNESE
GAAAGTGAGGCTGAGATTTTAAEILTGLT
ACTGGCCTCACGGTGGGCAGCVGSAADA
GCTGCAGATGCT
SS-022SecretionATGAAGCCTCTCCTTGTTGTG56MKPLLVV118
signalTTTGTCTTTCTTTTCCTTTGGGFVFLFLWD
ATCCAGTGCTGGCAPVLA
SS-023SecretionATGTCCTGTTCCCTAAAGTTT57MSCSLKFT119
signalACTTTGATTGTAATTTTTTTTTLIVIFFTCT
ACTGTTGGCTTTCATCCAGCLSSS
SS-024SecretionATGGTTCTTACTAAACCTCTTC58MVLTKPL120
signalAAAGAAATGGCAGCATGATGQRNGSMM
AGCTTTGAAAATGTGAAAGAASFENVKEK
AAGAGCAGAGAAGGAGGGCCSREGGPHA
CCATGCACACACACCCGAAGAHTPEEELC
AGAATTGTGTTTCGTGGTAACFVVTHTPQ
ACACTACCCTCAGGTTCAGACVQTTLNLF
CACACTCAACCTGTTTTTCCATFHIFKVLT
ATATTCAAGGTTCTTACTCAAQPLSLLWG
CCACTTTCCCTTCTGTGGGGT
SS-025SecretionATGGCCACCCCGCCATTCCGG59MATPPFRL121
signalCTGATAAGGAAGATGTTTTCCIRKMFSFK
TTCAAGGTGAGCAGATGGATGVSRWMGL
GGGCTTGCCTGCTTCCGGTCCACFRSLAAS
CTGGCGGCATCC
SS-026SecretionATGAGCTTTTTCCAACTCCTG60MSFFQLL122
signalATGAAAAGGAAGGAACTCATMKRKELIP
TCCCTTGGTGGTGTTCATGACLVVFMTV
TGTGGCGGCGGGTGGAGCCTCAAGGASS
ATCT
SS-027SecretionATGGTCTCAGCTCTGCGGGGA61MVSALRG123
signalGCACCCCTGATCAGGGTGCACAPLIRVHS
TCAAGCCCTGTTTCTTCTCCTTSPVSSPSV
CTGTGAGTGGACCACGGAGGCSGPAALVS
TGGTGAGCTGCCTGTCATCCCCLSSQSSA
AAAGCTCAGCTCTGAGCLS
SS-028SecretionATGATGGGGTCCCCAGTGAGT62MMGSPVS124
signalCATCTGCTGGCCGGCTTCTGTHLLAGFC
GTGTGGGTCGTCTTGGGCVWVVLG
SS-029SecretionATGGCAAGCATGGCTGCCGTG63MASMAAV125
signalCTCACCTGGGCTCTGGCTCTTLTWALAL
CTTTCAGCGTTTTCGGCCACCLSAFSATQA
CAGGCA
SS-030SecretionATGGTGCTCATGTGGACCAGT64MVLMWTS126
signalGGTGACGCCTTCAAGACGGCCGDAFKTA
TACTTCCTGCTGAAGGGTGCCYFLLKGAP
CCTCTGCAGTTCTCCGTGTGCLQFSVCGL
GGCCTGCTGCAGGTGCTGGTGLQVLVDL
GACCTGGCCATCCTGGGGCAGAILGQATA
GCCTACGCC
SS-031SecretionATGGATTTTGTCGCTGGAGCC65MDFVAGA127
signalATCGGAGGCGTCTGCGGTGTTIGGVCGV
GCTGTGGGCTACCCCCTGGACAVGYPLD
ACGGTGAAGGTCAGGATCCATVKVRIQT
GACGGAGCCAAAGTACACAGEPLYTGIW
GCATCTGGCACTGCGTCCGGGHCVRDTY
ATACGTATCACCGAGAGCGCGHRERVWG
TGTGGGFYRGLSLP
GCTTCTACCGGGGCCTCTCGCVCTVSLVSS
TGCCCGTGTGCACGGTGTCCC
TGGTATCTTCC
SS-032SecretionATGGAGAAGCCCCTCTTCCCA66MEKPLFPL128
signalTTAGTGCCTTTGCATTGGTTTGVPLHWFG
GCTTTGGCTACACAGCACTGGFGYTALV
TTGTTTCTGGTGGGATCGTTGVSGGIVGY
GCTATGTAAAAACAGGCAGCVKTGSVPS
GTGCCGTCCCTGGCTGCAGGGLAAGLLFG
CTGCTCTTCGGCAGTCTAGCCSLA
SS-033SecretionATGGGTCTGCTCCTTCCCCTG67MGLLLPL129
signalGCACTCTGCATCCTAGTCCTGALCILVLC
TGC
SS-034SecretionATGGGGATCCAGACGAGCCCC68MGIQTSPV130
signalGTCCTGCTGGCCTCCCTGGGGLLASLGVG
GTGGGGCTGGTCACTCTGCTCLVTLLGLA
GGCCTGGCTGTGGGCVG
SS-035SecretionATGTCGGACCTGCTACTACTG69MSDLLLL131
signalGGCCTGATTGGGGGCCTGACTGLIGGLTL
CTCTTACTGCTGCTGACGCTGLLLLTLLA
CTAGCCTTTGCCFA
SS-036SecretionATGGAGACTGTGGTGATTGTT70METVVIV132
signalGCCATAGGTGTGCTGGCCACCAIGVLATI
ATGTTTCTGGCTTCGTTTGCAGFLASFAAL
CCTTGGTGCTGGTTTGCAGGCVLVCRQ
AG
SS-037SecretionATGCGCGGCTCTGTGGAGTGC71MAGSVEC133
signalACCTGGGGTTGGGGGCACTGTTWGWGH
GCCCCCAGCCCCCTGCTCCTTCAPSPLLL
TGGACTCTACTTCTGTTTGCAWTLLLFA
GCCCCATTTGGCCTGCTGGGGAPFGLLG
SS-038SecretionATGATGCCGTCCCGTACCAAC72MMPSRTN134
signalCTGGCTACTGGAATCCCCAGTLATGIPSS
AGTAAAGTGAAATATTCAAGGKVKYSRLS
CTCTCCAGCACAGACGATGGCSTDDGYID
TACATTGACCTTCAGTTTAAGLQFKKTPP
AAAACCCCTCCTAAGATCCCTKIPYKAIA
TATAAGGCCATCGCACTTGCCLATVLFLI
ACTGTGCTGTTTTTGATTGGCGA
GCC
SS-039SecretionATGGCCCTGCCCCAGATGTGT73MALPQMC135
signalGACGGGAGCCACTTGGCCTCCDGSHLAST
ACCCTCCGCTATTGCATGACALRYCMTV
GTCAGCGGCACAGTGGTTCTGSGTVVLV
GTGGCCGGGACGCTCTGCTTCAGTLCFA
GCT
SS-041Vrg-6TGAAAAAGTGGTTCGTTGCTG74MKKWFVA136
CCGGCATCGGCGCTGCCGGACAGIGAGLL
TCATGCTCTCCAGCGCCGCCAMLSSAA
SS-042PhoAATGAAACAGAGCACCATTGCG75MKQSTIAL137
CTGGCGCTGCTGCCGCTGCTGALLPLLFT
TTTACCCCGGTGACCAAAGCGPVTKA
SS-043OmpAATGAAAAAAACCGCGATTGC76MKKTAIAI138
GATTGCGGTGGCGCTGGCGGGAVALAGF
CTTTGCGACCGTGGCGCAGGCGATVAQA
SS-044STIATGAAAAAACTGATGCTGGCG77MKKLMLA139
ATTTTTTTTAGCGTGCTGAGCTIFFSVLSFP
TTCCGAGCTTTAGCCAGAGCSFSQS
SS-045STIIATGAAAAAAAACATTGCGTTT78MKKNIAFL140
CTGCTGGCGAGCATGTTTGTGLASMFVFS
TTTAGCATTGCGACCAACGCGIATNAYA
TATGCG
SS-046AmylaseATGTTTGCGAAACGCTTTAAA79MFAKRFK141
ACCAGCCTGCTGCCGCTGTTTTSLLPLFA
GCGGGCTTTCTGCTGCTGTTTCGFLLLFHL
ATCTGGTGCTGGCGGGCCCGGVLAGPAA
CGGCGGCGAGCAS
SS-047AlphaATGCGCTTTCCGAGCATTTTT80MRFPSIFT142
FactorACCGCGGTGCTGTTTGCGGCGAVLFAASS
AGCAGCGCGCTGGCGALA
SS-048AlphaATGCGCTTTCCGAGCATTTTT81MRFPSIFT143
FactorACCACCGTGCTGTTTGCGGCGTVLFAASS
AGCAGCGCGCTGGCGALA
SS-049AlphaATGCGCTTTCCGAGCATTTTT82MRFPSIFTS144
FactorACCAGCGTGCTGTTTGCGGCGVLFAASSA
AGCAGCGCGCTGGCGLA
SS-050AlphaATGCGCTTTCCGAGCATTTTT83MRFPSIFT145
FactorACCCATGTGCTGTTTGCGGCGHVLFAASS
AGCAGCGCGCTGGCGALA
SS-051AlphaATGCGCTTTCCGAGCATTTTT84MRFPSIFTI146
FactorACCATTGTGCTGTTTGCGGCGVLFAASSA
AGCAGCGCGCTGGCGLA
SS-052AlphaATGCGCTTTCCGAGCATTTTT85MRFPSIFTF147
FactorACCTTTGTGCTGTTTGCGGCGVLFAASSA
AGCAGCGCGCTGGCGLA
SS-053AlphaATGCGCTTTCCGAGCATTTTT86MRFPSIFT148
FactorACCGAAGTGCTGTTTGCGGCGEVLFAASS
AGCAGCGCGCTGGCGALA
SS-054AlphaATGCGCTTTCCGAGCATTTTT87MRFPSIFT149
FactorACCGGCGTGCTGTTTGCGGCGGVLFAASS
AGCAGCGCGCTGGCGALA
SS-055Endoglucanase VATGCGTTCCTCCCCCCTCCTCC88MRSSPLLR150
GCTCCGCCGTTGTGGCCGCCCSAVVAAL
TGCCGGTGTTGGCCCTTGCCPVLALA
SS-056SecretionATGGGCGCGGCGGCCGTGCGC89MGAAAVR151
signalTGGCACTTGTGCGTGCTGCTGWHLCVLL
GCCCTGGGCACACGCGGGCGALGTRGRL
GCTG
SS-057FungalATGAGGAGCTCCCTTGTGCTG90MRSSLVLF152
TTCTTTGTCTCTGCGTGGACGFVSAWTA
GCCTTGGCCAGLA
SS-058FibronectinATGCTCAGGGGTCCGGGACCC91MLRGPGP153
GGGCGGCTGCTGCTGCTAGCAGRLLLLAV
GTCCTGTGCCTGGGGACATCGLCLGTSVR
GTGCGCTGCACCGAAACCGGGCTETGKSKR
AAGAGCAAGAGG
SS-059FibronectinATGCTTAGGGGTCCGGGGCCC92MLRGPGP154
GGGCTGCTGCTGCTGGCCGTCGLLLLAV
CAGCTGGGGACAGCGGTGCCCQCLGTAV
TCCACGPSTGA
SS-060FibronectinATGCGCCGGGGGGCCCTGACC93MRRGALT155
GGGCTGCTCCTGGTCCTGTGCGLLLVLCL
CTGAGTGTTGTGCTACGTGCASVVLRAAP
GCCCCCTCTGCAACAAGCAAGSATSKKRR
AAGCGCAGG

In the table, SS is secretion signal and MLS is mitochondrial leader signal. The signal-sensor primary constructs or mmRNA of the present invention may be designed to encode any of the signal peptide sequences of SEQ ID NOs 94-155, or fragments or variants thereof. These sequences may be included at the beginning of the oncology-related polypeptide coding region, in the middle or at the terminus or alternatively into a flanking region. Further, any of the signal-sensor polynucleotide primary constructs of the present invention may also comprise one or more of the sequences defined by SEQ ID NOs 32-93. These may be in the first region or either flanking region.

Additional signal peptide sequences which may be utilized in the present invention include those taught in, for example, databases such as those found at http://www.signalpeptide.de/ or http://proline.bic.nus.edu.sg/spdb/. Those described in U.S. Pat. Nos. 8,124,379; 7,413,875 and 7,385,034 are also within the scope of the invention and the contents of each are incorporated herein by reference in their entirety.

In one embodiment, the signal-sensor polynucleotide, primary constructs or mmRNA may include a nucleic acid sequence encoding a nuclear localization signal (NLS) and/or a nuclear export signal (NES). In one aspect, a signal-sensor polynucleotide, primary constructs or mmRNA may include a nucleic acid sequence encoding a nuclear localization signal (NLS). The signal-sensor polynucleotide, primary construct or mmRNA encoding a NLS would be able to traffic an oncology related polypeptide into the nucleus and deliver a survival or death signal to the nuclear microenvironment. In another aspect, the signal-sensor polynucleotide, primary constructs or mmRNA may include a nucleic acid sequence encoding a nuclear export signal such as NES 1 and/or NES2. As a nonlimiting example, the signal-sensor polynucleotide, primary constructs or mmRNA may encode a NES1, NES2 and a NLS signal and an oncology related polypeptide or a scambled sequence which is not translatable in order to interact with HIF1-alpha to alter the transcritome of the cancer cells.

Target Selection

According to the present invention, the signal-sensor primary constructs comprise at least a first region of linked nucleosides encoding at least one oncology-related polypeptide of interest. The oncology-related polypeptides of interest or “targets” or oncology-related proteins and oncology-related peptides of the present invention are listed in Table 6, Table 7 and Table 41. Oncology-related polypeptides may be divided into classes based on their function and area of cancer intervention. For example, the classes may include targets associated with (1) apoptosis or Survival signal imbalance (AS targets). These may be caspase dependent or caspase independent targets; (2) replicative potential or anti-senescence (CC/S targets); (3) metabolic stress including the involvement of acidosis or hypoxia (O2>1%) (M targets); (4) immune response (I targets); and (5) DNA damage/protection (DDR targets).

Shown in Table 6, in addition to the name and description of the gene encoding the oncology-related polypeptide of interest are the ENSEMBL Transcript ID (ENST), the ENSEMBL Protein ID (ENSP), each present where applicable, and when available the optimized sequence ID (OPT. SEQ ID). The targets are also categorized by group where “AS” refers to targets involved in apoptotic signaling; “M” refers to targets involved in metabolic processes and “CC/S” refers to targets involved in cell cycle and senescense.

TABLE 6
Oncology Related Targets
Prot.OPT.
Trans.SEQSEQ
ENSTSEQENSPIDID
Cat.TargetTarget DescriptionIDID NOIDNONO
AS14-3-3tyrosine 3-2380811562380811321
monooxygenase/tryptophan 5-
monooxygenase activation
protein, theta polypeptide
AS14-3-3tyrosine 3-2489751572489751322
monooxygenase/tryptophan 5-
monooxygenase activation
protein, eta polypeptide
AS14-3-3tyrosine 3-2643351582643351323
monooxygenase/tryptophan 5-
monooxygenase activation
protein, epsilon polypeptide
AS14-3-3tyrosine 3-3076301593063301324
monooxygenase/tryptophan 5-
monooxygenase activation
protein, gamma polypeptide
AS14-3-3tyrosine 3-3532451603095031325
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-3537031613001611326
monooxygenase/tryptophan 5-
monooxygenase activation
protein, beta polypeptide
AS14-3-3tyrosine 3-3728391623619301327
monooxygenase/tryptophan 5-
monooxygenase activation
protein, beta polypeptide
AS14-3-3tyrosine 3-3818441633712671328
monooxygenase/tryptophan 5-
monooxygenase activation
protein, theta polypeptide
AS14-3-3tyrosine 3-3959481643792781329
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-3959511653792811330
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-3959531663792831331
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-3959561673792861332
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-3959571683792871333
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-3959581693792881334
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-4141311704060581335
monooxygenase/tryptophan 5-
monooxygenase activation
protein, epsilon polypeptide
AS14-3-3tyrosine 3-4189971714165511336
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-4194771723951141337
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-4282621733947291338
monooxygenase/tryptophan 5-
monooxygenase activation
protein, beta polypeptide
AS14-3-3tyrosine 3-4372931743948801339
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-4458301753945581340
monooxygenase/tryptophan 5-
monooxygenase activation
protein, beta polypeptide
AS14-3-3tyrosine 3-4466191763989901341
monooxygenase/tryptophan 5-
monooxygenase activation
protein, theta polypeptide
AS14-3-3tyrosine 3-4532071773906451342
monooxygenase/tryptophan 5-
monooxygenase activation
protein, gamma polypeptide
AS14-3-3tyrosine 3-4573091783985991343
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-5177971794278011344
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-5213091804296231345
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-5213281814290411346
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-5216071824300581347
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-5225421834300721348
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-5228191844287751349
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-5231311854283811350
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-5238481864288601351
monooxygenase/tryptophan 5-
monooxygenase activation
protein, zeta polypeptide
AS14-3-3tyrosine 3-5367551874438031352
monooxygenase/tryptophan 5-
monooxygenase activation
protein, gamma polypeptide
AS14-3-3tyrosine 3-5399791884432261353
monooxygenase/tryptophan 5-
monooxygenase activation
protein, theta polypeptide
ASAIFapoptosis-inducing factor,2872951892872951354
mitochondrion-associated, 1
ASAIFapoptosis-inducing factor,3078641903123701355
mitochondrion-associated, 2
ASAIFapoptosis-inducing factor,3199081913151221356
mitochondrion-associated, 1
ASAIFapoptosis-inducing factor,3336071923276711357
mitochondrion-associated, 3
ASAIFapoptosis-inducing factor,3353751933353691358
mitochondrion-associated, 3
ASAIFapoptosis-inducing factor,3464241943163201359
mitochondrion-associated, 1
ASAIFapoptosis-inducing factor,3732481953623451360
mitochondrion-associated, 2
ASAIFapoptosis-inducing factor,3950391963784801361
mitochondrion-associated, 2
ASAIFapoptosis-inducing factor,3991631973821161362
mitochondrion-associated, 3
ASAIFapoptosis-inducing factor,3991671983821201363
mitochondrion-associated, 3
ASAIFapoptosis-inducing factor,4050891993858001364
mitochondrion-associated, 3
ASAIFapoptosis-inducing factor,4347142003996571365
mitochondrion-associated, 3
ASAIFapoptosis-inducing factor,4402382013907981366
mitochondrion-associated, 3
ASAIFapoptosis-inducing factor,4402632024058791367
mitochondrion-associated, 1
ASAIFapoptosis-inducing factor,4413762034020671368
mitochondrion-associated, 3
ASAIFapoptosis-inducing factor,4604362044312221369
mitochondrion-associated, 1
ASAIFapoptosis-inducing factor,5357242054461131370
mitochondrion-associated, 1
ASAKTv-akt murine thymoma viral2638262062638261371
(PKB)oncogene homolog 3 (protein
kinase B, gamma)
ASAKTv-akt murine thymoma viral3112782073094281372
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral3361992083369431373
(PKB)oncogene homolog 3 (protein
kinase B, gamma)
ASAKTv-akt murine thymoma viral3493102092702021374
(PKB)oncogene homolog 1
ASAKTv-akt murine thymoma viral3583352103510951375
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral3665392113554971376
(PKB)oncogene homolog 3 (protein
kinase B, gamma)
ASAKTv-akt murine thymoma viral3665402123554981377
(PKB)oncogene homolog 3 (protein
kinase B, gamma)
ASAKTv-akt murine thymoma viral3918442133757191378
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral3920372143758911379
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral3920382153758921380
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral4026152163853261381
(PKB)oncogene homolog 1
ASAKTv-akt murine thymoma viral4077962173842931382
(PKB)oncogene homolog 1
ASAKTv-akt murine thymoma viral4163622184079991383
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral4169942193924581384
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral4231272204038421385
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral4249012213995321386
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral4273752224038901387
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral4520772234040831388
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral4564412243965321389
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral5378342254415911390
(PKB)oncogene homolog 2
ASAKTv-akt murine thymoma viral5441682264438971391
(PKB)oncogene homolog 1
ASAKTv-akt murine thymoma viral5526312274478201392
(PKB)oncogene homolog 3 (protein
kinase B, gamma)
ASAKTv-akt murine thymoma viral5545812284518281393
(PKB)oncogene homolog 1
ASAKTv-akt murine thymoma viral5548482294511661394
(PKB)oncogene homolog 1
ASAKTv-akt murine thymoma viral5555282304506881395
(PKB)oncogene homolog 1
ASAKTv-akt murine thymoma viral5559262314518241396
(PKB)oncogene homolog 1
ASANTsolute carrier family 252814562322814561397
(mitochondrial carrier; adenine
nucleotide translocator),
member 4
ASApaf-1apoptotic peptidase activating3339912333345581398
factor 1
ASApaf-1apoptotic peptidase activating3394332343418301399
factor 1
ASApaf-1apoptotic peptidase activating3573102353498621400
factor 1
ASApaf-1apoptotic peptidase activating3599722363530591401
factor 1
ASApaf-1apoptotic peptidase activating5470452374497911402
factor 1
ASApaf-1apoptotic peptidase activating5490072384481611403
factor 1
ASApaf-1apoptotic peptidase activating5505272394484491404
factor 1
ASApaf-1apoptotic peptidase activating5519642404481651405
factor 1
ASApaf-1apoptotic peptidase activating5522682414488261406
factor 1
ASAPRILtumor necrosis factor (ligand)3387842423435051407
(TNFSF13)superfamily, member 13
ASAPRILtumor necrosis factor (ligand)3492282433144551408
(TNFSF13)superfamily, member 13
ASAPRILtumor necrosis factor (ligand)3805352443699081409
(TNFSF13)superfamily, member 13
ASAPRILtumor necrosis factor (ligand)3965452453797941410
(TNFSF13)superfamily, member 13
ASARTSphosphoribosyl pyrophosphate3724182463614951411
synthetase 1
ASARTSphosphoribosyl pyrophosphate3724192473614961412
synthetase 1
ASARTSphosphoribosyl pyrophosphate3724282483615051413
synthetase 1
ASARTSphosphoribosyl pyrophosphate3724352493615121414
synthetase 1
ASARTSphosphoribosyl pyrophosphate5432482504431851415
synthetase 1
ASASK1mitogen-activated protein3558452513481041416
(MAP3K5)kinase kinase kinase 5
ASASK1mitogen-activated protein3590152523519081417
(MAP3K5)kinase kinase kinase 5
ASASK1mitogen-activated protein3677682533567421418
(MAP3K5)kinase kinase kinase 5
ASBADBCL2-associated agonist of3090322543091031419
cell death
ASBADBCL2-associated agonist of3945322553780401420
cell death
ASBADBCL2-associated agonist of5401522564408071421
cell death
ASBAFF(TNFSF13B)tumor necrosis factor (ligand)3758872573650481422
superfamily, member 13b
ASBAFF(TNFSF13B)tumor necrosis factor (ligand)4305592583895401423
superfamily, member 13b
ASBAFF(TNFSF13B)tumor necrosis factor (ligand)5421362594453341424
superfamily, member 13b
ASBakBCL2-antagonist/killer 13606612603538781425
ASBakBCL2-antagonist/killer 13744602613635841426
ASBakBCL2-antagonist/killer 13744672623635911427
ASBakBCL2-antagonist/killer 14429982633912581428
ASBAXBCL2-associated X protein2932882642932881429
ASBAXBCL2-associated X protein3453582652632621430
ASBAXBCL2-associated X protein3544702663464611431
ASBAXBCL2-associated X protein3918712673757441432
ASBAXBCL2-associated X protein4159692683899711433
ASBAXBCL2-associated X protein5397872694414131434
ASBcl-2B-cell CLL/lymphoma 23336812703296231435
ASBcl-2B-cell CLL/lymphoma 23981172713811851436
ASBcl-2B-cell CLL/lymphoma 24444842724042141437
ASBcl-BBCL2-like 10 (apoptosis2604422732604421438
facilitator)
ASBcl-WBCL2-like 22504052742504051439
ASBcl-WBCL2-like 25546352754512341440
ASBcl-WBCL2-like 25572362764517011441
ASBcl-WBCL2-like 25575792774522651442
ASBcl-XLBCL2-like 13076772783025641443
ASBcl-XLBCL2-like 13760552793652231444
ASBcl-XLBCL2-like 13760622803652301445
ASBcl-XLBCL2-like 14204882813907601446
ASBcl-XLBCL2-like 14206532824055631447
ASBcl-XLBCL2-like 14229202834112521448
ASBcl-XLBCL2-like 14392672843896881449
ASBcl-XLBCL2-like 14502732854062031450
ASBcl-XLBCL2-like 14564042863955451451
ASBCMAtumor necrosis factor receptor 53243287 532431452
superfamily, member 17
ASBCMAtumor necrosis factor receptor3964952883797531453
superfamily, member 17
ASBCMAtumor necrosis factor receptor4353552894017821454
superfamily, member 17
ASBFL1BCL2-related protein A12679532902679531455
ASBFL1BCL2-related protein A13356612913352501456
ASBidBH3 interacting domain death3173612923188221457
agonist
ASBidBH3 interacting domain death3421112933445941458
agonist
ASBidBH3 interacting domain death3997652943826671459
agonist
ASBidBH3 interacting domain death3997672953826691460
agonist
ASBidBH3 interacting domain death3997742963826741461
agonist
ASBidBH3 interacting domain death5519522974492361462
agonist
ASBikBCL2-interacting killer2161152982161151463
(apoptosis-inducing)
ASBimBCL2-like 11 (apoptosis3086592993092261464
facilitator)
ASBimBCL2-like 11 (apoptosis3375653003383741465
facilitator)
ASBimBCL2-like 11 (apoptosis3577573013503981466
facilitator)
ASBimBCL2-like 11 (apoptosis3932523023769411467
facilitator)
ASBimBCL2-like 11 (apoptosis3932533033769421468
facilitator)
ASBimBCL2-like 11 (apoptosis3932563043769431469
facilitator)
ASBimBCL2-like 11 (apoptosis4321793054118701470
facilitator)
ASBimBCL2-like 11 (apoptosis4520333064036661471
facilitator)
ASBMFBcl2 modifying factor2204463072204461472
ASBMFBcl2 modifying factor3546703083466971473
ASBMFBcl2 modifying factor3975733093807031474
ASBMFBcl2 modifying factor4314153103965111475
ASBMFBcl2 modifying factor5597013114539191476
ASBMFBcl2 modifying factor5612823124535221477
ASBMFBcl2 modifying factor5613603134538921478
ASBREbrain and reproductive organ-3420453143393711479
expressed (TNFRSF1A
modulator)
ASBREbrain and reproductive organ-3447733153434121480
expressed (TNFRSF1A
modulator)
ASBREbrain and reproductive organ-3617043163546991481
expressed (TNFRSF1A
modulator)
ASBREbrain and reproductive organ-3796233173689441482
expressed (TNFRSF1A
modulator)
ASBREbrain and reproductive organ-3796243183689451483
expressed (TNFRSF1A
modulator)
ASBREbrain and reproductive organ-3796323193689531484
expressed (TNFRSF1A
modulator)
ASBREbrain and reproductive organ-4369243203923451485
expressed (TNFRSF1A
modulator)
ASCalcineurin Aprotein phosphatase 3, catalytic3230553213205801486
subunit, alpha isozyme
ASCalcineurin Aprotein phosphatase 3, catalytic3948533223783221487
subunit, alpha isozyme
ASCalcineurin Aprotein phosphatase 3, catalytic3948543233783231488
subunit, alpha isozyme
ASCalcineurin Aprotein phosphatase 3, catalytic5071763244229901489
subunit, alpha isozyme
ASCalcineurin Aprotein phosphatase 3, catalytic5122153254227811490
subunit, alpha isozyme
ASCalcineurin Aprotein phosphatase 3, catalytic5236943264293501491
subunit, alpha isozyme
ASCalcineurin Aprotein phosphatase 3, catalytic5258193274345991492
subunit, alpha isozyme
ASCalcineurin Aprotein phosphatase 3, catalytic5293243284316191493
subunit, alpha isozyme
ASCaspase-1caspase 1, apoptosis-related3532473293441321494
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related3931363303768441495
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related4159813314084461496
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related4368633324100761497
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related4463693334032601498
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related5258253344347791499
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related5265683354342501500
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related5289743364342591501
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related5298713374319471502
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related5311663384343031503
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related5334003394331381504
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-1caspase 1, apoptosis-related5344973404368751505
cysteine peptidase (interleukin
1, beta, convertase)
ASCaspase-caspase 10, apoptosis-related2728793412728791506
10cysteine peptidase
ASCaspase-caspase 10, apoptosis-related2861863422861861507
10cysteine peptidase
ASCaspase-caspase 10, apoptosis-related3468173432378651508
10cysteine peptidase
ASCaspase-caspase 10, apoptosis-related3601323443532501509
10cysteine peptidase
ASCaspase-2caspase 2, apoptosis-related3104473453126641510
cysteine peptidase
ASCaspase-2caspase 2, apoptosis-related3506233463400301511
cysteine peptidase
ASCaspase-2caspase 2, apoptosis-related3929233473766541512
cysteine peptidase
ASCaspase-3caspase 3, apoptosis-related3083943483110321513
cysteine peptidase
ASCaspase-3caspase 3, apoptosis-related4384673493907921514
cysteine peptidase
ASCaspase-3caspase 3, apoptosis-related4471213504071421515
cysteine peptidase
ASCaspase-3caspase 3, apoptosis-related5239163514289291516
cysteine peptidase
ASCaspase-4caspase 4, apoptosis-related3555463523477411517
cysteine peptidase
ASCaspase-4caspase 4, apoptosis-related4174403534016731518
cysteine peptidase
ASCaspase-4caspase 4, apoptosis-related4447393543885661519
cysteine peptidase
ASCaspase-5caspase 5, apoptosis-related2603153552603151520
cysteine peptidase
ASCaspase-5caspase 5, apoptosis-related3931393563768471521
cysteine peptidase
ASCaspase-5caspase 5, apoptosis-related3931413573768491522
cysteine peptidase
ASCaspase-5caspase 5, apoptosis-related4184343583981301523
cysteine peptidase
ASCaspase-5caspase 5, apoptosis-related4447493593883651524
cysteine peptidase
ASCaspase-5caspase 5, apoptosis-related5260563604368771525
cysteine peptidase
ASCaspase-5caspase 5, apoptosis-related5313673614344711526
cysteine peptidase
ASCaspase-6caspase 6, apoptosis-related2651643622651641527
cysteine peptidase
ASCaspase-6caspase 6, apoptosis-related3529813632853331528
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related3456333642987011529
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related3693153653583211530
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related3693163663583221531
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related3693183673583241532
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related3693193683583251533
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related3693213693583271534
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related3693313703583371535
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related4296173714000941536
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related4423933723944821537
cysteine peptidase
ASCaspase-7caspase 7, apoptosis-related4524903733981071538
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related2642743742642741539
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related2642753752642751540
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related3234923763257221541
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related3584853773512731542
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related3922583783760871543
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related3922593793760881544
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related3922613803760891545
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related3922633813760911546
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related3922663823760941547
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related4137263833975281548
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related4298813843906411549
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related4321093854125231550
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related4407323863968691551
cysteine peptidase
ASCaspase-8caspase 8, apoptosis-related4476163873883061552
cysteine peptidase
ASCaspase-9caspase 9, apoptosis-related3338683883302371553
cysteine peptidase
ASCaspase-9caspase 9, apoptosis-related3485493892552561554
cysteine peptidase
ASCaspase-9caspase 9, apoptosis-related3758743903650341555
cysteine peptidase
ASCaspase-9caspase 9, apoptosis-related3758903913650511556
cysteine peptidase
ASCaspase-9caspase 9, apoptosis-related4404843924113041557
cysteine peptidase
ASCaspase-9caspase 9, apoptosis-related4475223933965401558
cysteine peptidase
ASCaspase-9caspase 9, apoptosis-related5464243944495841559
cysteine peptidase
ASCD27CD27 molecule2665573952665571560
ASCD30tumor necrosis factor receptor2639323962639321561
superfamily, member 8
ASCD30tumor necrosis factor receptor4131463973983371562
superfamily, member 8
ASCD30tumor necrosis factor receptor4178143983906501563
superfamily, member 8
ASCD30Ltumor necrosis factor (ligand)2237953992237951564
superfamily, member 8
ASCD40CD40 molecule, TNF receptor3722784003613521565
superfamily member 5
ASCD40LCD40 ligand3706284013596621566
(TNFSF5)
ASCD40LCD40 ligand3706294023596631567
(TNFSF5)
ASCD41CD40 molecule, TNF receptor3722764033613501568
superfamily member 5
ASCD42CD40 molecule, TNF receptor3722854043613591569
superfamily member 5
ASCD70(TNFSF7)CD70 molecule2459034052459031570
ASCD70(TNFSF7)CD70 molecule4231454063952941571
ASCDK1cyclin-dependent kinase 13166294073259701572
(p34)
ASCDK1cyclin-dependent kinase 13738094083629151573
(p34)
ASCDK1cyclin-dependent kinase 13952844093786991574
(p34)
ASCDK1cyclin-dependent kinase 14482574103979731575
(p34)
ASCDK1cyclin-dependent kinase 15190784114306651576
(p34)
ASCDK5cyclin-dependent kinase 54859724124197821577
ASCDK5R1cyclin-dependent kinase 5,3134014133184861578
(p35)regulatory subunit 1 (p35)
ASc-CASP8 and FADD-like3099554143124551579
FLIP(S)apoptosis regulator
ASc-CASP8 and FADD-like3408704153393261580
FLIP(S)apoptosis regulator
ASc-CASP8 and FADD-like3433754163393911581
FLIP(S)apoptosis regulator
ASc-CASP8 and FADD-like3555584173477571582
FLIP(S)apoptosis regulator
ASc-CASP8 and FADD-like3951484183785801583
FLIP(S)apoptosis regulator
ASc-CASP8 and FADD-like4177484194128821584
FLIP(S)apoptosis regulator
ASc-CASP8 and FADD-like4232414203994201585
FLIP(S)apoptosis regulator
ASc-CASP8 and FADD-like4334454213910291586
FLIP(S)apoptosis regulator
ASc-CASP8 and FADD-like4412244224118971587
FLIP(S)apoptosis regulator
ASc-CASP8 and FADD-like4432274234132701588
FLIP(S)apoptosis regulator
AScIAP1baculoviral IAP repeatNA424NA15892488
containing 3
ASc-IAP1baculoviral IAP repeat2634644252634641590
containing 3
ASc-IAP1baculoviral IAP repeat5328084264329071591
containing 3
AScIAP2baculoviral IAP repeatNA427NA1592
containing 2
ASC-IAP2baculoviral IAP repeat2277584282277581593
containing 2
ASC-IAP2baculoviral IAP repeat5306754294317231594
containing 2
ASC-IAP2baculoviral IAP repeat5326724304349791595
containing 2
ASC-IAP2baculoviral IAP repeat5417414314407711596
containing 2
ASc-Junjun proto-oncogene3712224323602661597
ASc-Raf-1v-raf-1 murine leukemia viral2518494332518491598
oncogene homolog 1
ASc-Raf-1v-raf-1 murine leukemia viral4424154344018881599
oncogene homolog 1
ASc-Raf-1v-raf-1 murine leukemia viral5349974354411861600
oncogene homolog 1
ASc-Raf-1v-raf-1 murine leukemia viral5421774364435671601
oncogene homolog 1
ASCytochrome ccytochrome c, somatic3057864373077861602
ASCytochrome ccytochrome c, somatic4094094383862701603
ASCytochrome ccytochrome c, somatic4097644393872791604
ASCytochrome ccytochrome c, somatic4134474404164791605
ASDAXXdeath-domain associated2660004412660001606
protein
ASDAXXdeath-domain associated3745424423636681607
protein
ASDAXXdeath-domain associated3830624433725391608
protein
ASDAXXdeath-domain associated3831944443726811609
protein
ASDAXXdeath-domain associated3990604453820141610
protein
ASDAXXdeath-domain associated3993444463822811611
protein
ASDAXXdeath-domain associated4140834473968761612
protein
ASDAXXdeath-domain associated4142724484097561613
protein
ASDAXXdeath-domain associated4198554493976121614
protein
ASDAXXdeath-domain associated4282684504082151615
protein
ASDAXXdeath-domain associated4295314514158981616
protein
ASDAXXdeath-domain associated4334824524046231617
protein
ASDAXXdeath-domain associated4363114534043761618
protein
ASDAXXdeath-domain associated4383324544117001619
protein
ASDAXXdeath-domain associated4405004554039861620
protein
ASDAXXdeath-domain associated4450094563941081621
protein
ASDAXXdeath-domain associated4464034574060081622
protein
ASDAXXdeath-domain associated4534074584084991623
protein
ASDAXXdeath-domain associated4539314594124331624
protein
ASDAXXdeath-domain associated4541974604121771625
protein
ASDAXXdeath-domain associated4558604614107721626
protein
ASDAXXdeath-domain associated5476634624471151627
protein
ASDAXXdeath-domain associated5486044634483371628
protein
ASDAXXdeath-domain associated5508224644478611629
protein
ASDAXXdeath-domain associated5529444654478331630
protein
ASDcR3tumor necrosis factor receptor3428524663423281631
superfamily, member 6b,
decoy
ASDcR3tumor necrosis factor receptor3699964673590131632
superfamily, member 6b,
decoy
ASDcR3tumor necrosis factor receptor3700064683590231633
superfamily, member 6b,
decoy
ASDFF40DNA fragmentation factor,3388954693395241634
(CAD)40 kDa, beta polypeptide
(caspase-activated DNase)
ASDFF40DNA fragmentation factor,3393504703432181635
(CAD)40 kDa, beta polypeptide
(caspase-activated DNase)
ASDFF40DNA fragmentation factor,3413854713459061636
(CAD)40 kDa, beta polypeptide
(caspase-activated DNase)
ASDFF40DNA fragmentation factor,3782064723674481637
(CAD)40 kDa, beta polypeptide
(caspase-activated DNase)
ASDFF40DNA fragmentation factor,3782094733674541638
(CAD)40 kDa, beta polypeptide
(caspase-activated DNase)
ASDFF40DNA fragmentation factor,3782124743674571639
(CAD)40 kDa, beta polypeptide
(caspase-activated DNase)
ASDFF40DNA fragmentation factor,4305394753895021640
(CAD)40 kDa, beta polypeptide
(caspase-activated DNase)
ASDFF40DNA fragmentation factor,4486324764116351641
(CAD)40 kDa, beta polypeptide
(caspase-activated DNase)
ASDFF40DNA fragmentation factor,4919984774367751642
(CAD)40 kDa, beta polypeptide
(caspase-activated DNase)
ASDR3tumor necrosis factor receptor3483334783144511643
superfamily, member 25
ASDR3tumor necrosis factor receptor3517484793267621644
superfamily, member 25
ASDR3tumor necrosis factor receptor3519594803377131645
superfamily, member 25
ASDR3tumor necrosis factor receptor3568764813493411646
superfamily, member 25
ASDR3tumor necrosis factor receptor3777824823670131647
superfamily, member 25
ASDR4tumor necrosis factor receptor2211324832211321648
superfamily, member 10a
ASDR5tumor necrosis factor receptor2764314842764311649
superfamily, member 10b
ASDR5tumor necrosis factor receptor3477394853178591650
superfamily, member 10b
ASDR5tumor necrosis factor receptor5422264864433861651
superfamily, member 10b
ASDR6tumor necrosis factor receptor2968614872968611652
superfamily, member 21
ASDR6tumor necrosis factor receptor4192064883900321653
superfamily, member 21
ASEGFRepidermal growth factor2754934892754931654
receptor
ASEGFRepidermal growth factor3429164903423761655
receptor
ASEGFRepidermal growth factor3445764913459731656
receptor
ASEGFRepidermal growth factor3955044923788801657
receptor
ASEGFRepidermal growth factor4203164934138431658
receptor
ASEGFRepidermal growth factor4425914944100311659
receptor
ASEGFRepidermal growth factor4547574953952431660
receptor
ASEGFRepidermal growth factor4550894964155591661
receptor
ASEGFRepidermal growth factor5334504974352621662
receptor
ASErbB2v-erb-b2 erythroblastic2695714982695711663
leukemia viral oncogene
homolog 2, neuro/glioblastoma
derived oncogene homolog
(avian)
ASErbB2v-erb-b2 erythroblastic4063814993851851664
leukemia viral oncogene
homolog 2, neuro/glioblastoma
derived oncogene homolog
(avian)
ASErbB2v-erb-b2 erythroblastic4456585004040471665
leukemia viral oncogene
homolog 2, neuro/glioblastoma
derived oncogene homolog
(avian)
ASErbB2v-erb-b2 erythroblastic5400425014463821666
leukemia viral oncogene
homolog 2, neuro/glioblastoma
derived oncogene homolog
(avian)
ASErbB2v-erb-b2 erythroblastic5401475024435621667
leukemia viral oncogene
homolog 2, neuro/glioblastoma
derived oncogene homolog
(avian)
ASErbB2v-erb-b2 erythroblastic5417745034464661668
leukemia viral oncogene
homolog 2, neuro/glioblastoma
derived oncogene homolog
(avian)
ASErbB3v-erb-b2 erythroblastic2671015042671011669
leukemia viral oncogene
homolog 3 (avian)
ASErbB3v-erb-b2 erythroblastic3940995053776591670
leukemia viral oncogene
homolog 3 (avian)
ASErbB3v-erb-b2 erythroblastic4117315064157531671
leukemia viral oncogene
homolog 3 (avian)
ASErbB3v-erb-b2 erythroblastic4152885074083401672
leukemia viral oncogene
homolog 3 (avian)
ASErbB3v-erb-b2 erythroblastic4501465083991781673
leukemia viral oncogene
homolog 3 (avian)
ASErbB3v-erb-b2 erythroblastic5492825094486361674
leukemia viral oncogene
homolog 3 (avian)
ASErbB3v-erb-b2 erythroblastic5510855104484831675
leukemia viral oncogene
homolog 3 (avian)
ASErk(MAPK1/mitogen-activated protein2158325112158321676
3)kinase 1
ASErk(MAPK1/mitogen-activated protein2630255122630251677
3)kinase 3
ASErk(MAPK1/mitogen-activated protein3222665133272931678
3)kinase 3
ASErk(MAPK1/mitogen-activated protein3952005143786261679
3)kinase 3
ASErk(MAPK1/mitogen-activated protein3952025153786281680
3)kinase 3
ASErk(MAPK1/mitogen-activated protein3988225163818031681
3)kinase 1
ASErk(MAPK1/mitogen-activated protein4033945173848951682
3)kinase 3
ASErk(MAPK1/mitogen-activated protein4159115184091491683
3)kinase 1
ASErk(MAPK1/mitogen-activated protein4846635194327421684
3)kinase 3
ASErk(MAPK1/mitogen-activated protein5447865204408421685
3)kinase 1
ASFADDFas (TNFRSF6)-associated via3018385213018381686
death domain
ASFLASHcaspase 8 associated protein 2237177522NA1687
ASFLASHcaspase 8 associated protein 2419040523NA
ASFLASHcaspase 8 associated protein 2444163524NA
ASFLASHcaspase 8 associated protein 2547893525NA
ASFLASHcaspase 8 associated protein 2548224526NA
ASFLASHcaspase 8 associated protein 2551025527NA
ASFLASHcaspase 8 associated protein 2552401528NA
ASFN14tumor necrosis factor receptor3265775293267371688
superfamily, member 12A
ASFN14tumor necrosis factor receptor3416275303438941689
superfamily, member 12A
ASGCKmitogen-activated protein2940665312940661690
(MAP4K2)kinase kinase kinase kinase 2
ASGRB2growth factor receptor-bound3166155323173601691
protein 2
ASGRB2growth factor receptor-bound3168045333390071692
protein 2
ASGRB2growth factor receptor-bound3925625343763451693
protein 2
ASGRB2growth factor receptor-bound3925645353763471694
protein 2
ASH-Rasv-Ha-ras Harvey rat sarcoma3111895363098451695
viral oncogene homolog
ASH-Rasv-Ha-ras Harvey rat sarcoma3975945373807221696
viral oncogene homolog
ASH-Rasv-Ha-ras Harvey rat sarcoma3975965383807231697
viral oncogene homolog
ASH-Rasv-Ha-ras Harvey rat sarcoma4173025393882461698
viral oncogene homolog
ASH-Rasv-Ha-ras Harvey rat sarcoma4515905404075861699
viral oncogene homolog
ASH-Rasv-Ha-ras Harvey rat sarcoma4932305414340231700
viral oncogene homolog
ASHRKharakiri, BCL2 interacting2575725422575721701
protein (contains only BH3
domain)
ASHSP27heat shock 27 kDa protein 12485535432485531702
ASHSP27heat shock 27 kDa protein 33020055443033941703
ASHSP27Heat shock protein beta-23042985453024761704
ASHSP27heat shock 27 kDa protein 14322765464065451705
ASHSP27Heat shock protein beta-25373825474455851706
ASHtrA2/OmiHtrA serine peptidase 22580805482580801707
ASHtrA2/OmiHtrA serine peptidase 23522225493128931708
ASHumaninMT-RNR2-like 43999745503828561709
ASHumaninMT-RNR2-like 55125245514379101710
ASHumaninMT-RNR2-like 85366845524396661711
ASHumaninMT-RNR2-like 15400405534392281712
ASHumaninMT-RNR2-like 35435005544433391713
ASHumaninMT-RNR2-like 75448245554399851714
ASHumaninMT-RNR2-like 105450755564421591715
ASHumaninMT-RNR2-like 65704195574610751716
ASICADDNA fragmentation factor,3770365583662351717
45 kDa, alpha polypeptide
ASICADDNA fragmentation factor,3770385593662371718
45 kDa, alpha polypeptide
ASIGF-1Rinsulin-like growth factor 12680355602680351719
receptor
ASIKKconserved helix-loop-helix3703975613594241720
(alpha)ubiquitous kinase
ASIKKinhibitor of kappa light3797085623690301721
(beta)polypeptide gene enhancer in
B-cells, kinase beta
ASIKKinhibitor of kappa light4165055634049201722
(beta)polypeptide gene enhancer in
B-cells, kinase beta
ASIKKinhibitor of kappa light5208105644306841723
(beta)polypeptide gene enhancer in
B-cells, kinase beta
ASIKK-inhibitor of kappa light2635185652635181724
gammapolypeptide gene enhancer in
B-cells, kinase gamma
ASIKK-inhibitor of kappa light3696015663586141725
gammapolypeptide gene enhancer in
B-cells, kinase gamma
ASIKK-inhibitor of kappa light3696065673586191726
gammapolypeptide gene enhancer in
B-cells, kinase gamma
ASIKK-inhibitor of kappa light3696075683586201727
gammapolypeptide gene enhancer in
B-cells, kinase gamma
ASIKK-inhibitor of kappa light3696095693586221728
gammapolypeptide gene enhancer in
B-cells, kinase gamma
ASIKK-inhibitor of kappa light4226805703903681729
gammapolypeptide gene enhancer in
B-cells, kinase gamma
ASIKK-inhibitor of kappa light4402865713949341730
gammapolypeptide gene enhancer in
B-cells, kinase gamma
ASIKK-inhibitor of kappa light4456225723952051731
gammapolypeptide gene enhancer in
B-cells, kinase gamma
ASIKK-inhibitor of kappa light4555885734007691732
gammapolypeptide gene enhancer in
B-cells, kinase gamma
ASIRAK1interleukin-1 receptor-3699805743589971733
associated kinase 1
ASIRAK1interleukin-1 receptor-3936825753772871734
associated kinase 1
ASIRAK1interleukin-1 receptor-3936875763772911735
associated kinase 1
ASIRAK1interleukin-1 receptor-4299365773926621736
associated kinase 1
ASIRAK2interleukin-1 receptor-2564585782564581737
associated kinase 2
ASIRS-1insulin receptor substrate 13051235793048951738
ASjBid;jBIDNANA1739
formed
after
cleaving
BID at
position
25
ASJNK1(MAPK8)mitogen-activated protein3603325803534831740
kinase 8
ASJNK1(MAPK8)mitogen-activated protein3741745813632891741
kinase 8
ASJNK1(MAPK8)mitogen-activated protein3741765823632911742
kinase 8
ASJNK1(MAPK8)mitogen-activated protein3741795833632941743
kinase 8
ASJNK1(MAPK8)mitogen-activated protein3741825843632971744
kinase 8
ASJNK1(MAPK8)mitogen-activated protein3741895853633041745
kinase 8
ASJNK1(MAPK8)mitogen-activated protein3956115863789741746
kinase 8
ASJNK1(MAPK8)mitogen-activated protein4265575873977291747
kinase 8
ASJNK1(MAPK8)mitogen-activated protein4290415883932231748
kinase 8
ASJNK1(MAPK8)mitogen-activated protein4323795893879361749
kinase 8
ASJNK3(MAPK10)mitogen-activated protein3592215903521571750
kinase 10
ASJNK3(MAPK10)mitogen-activated protein3615695913552971751
kinase 10
ASJNK3(MAPK10)mitogen-activated protein3951575923785861752
kinase 10
ASJNK3(MAPK10)mitogen-activated protein3951605933785891753
kinase 10
ASJNK3(MAPK10)mitogen-activated protein3951615943785901754
kinase 10
ASJNK3(MAPK10)mitogen-activated protein3951665953785951755
kinase 10
ASJNK3(MAPK10)mitogen-activated protein3951695963785981756
kinase 10
ASJNK3(MAPK10)mitogen-activated protein4490475974144691757
kinase 10
ASJNK3(MAPK10)mitogen-activated protein5023025984239181758
kinase 10
ASJNK3(MAPK10)mitogen-activated protein5039115994214091759
kinase 10
ASJNK3(MAPK10)mitogen-activated protein5067736004213591760
kinase 10
ASJNK3(MAPK10)mitogen-activated protein5094646014241281761
kinase 10
ASJNK3(MAPK10)mitogen-activated protein5111676024222771762
kinase 10
ASJNK3(MAPK10)mitogen-activated protein5113286034217621763
kinase 10
ASJNK3(MAPK10)mitogen-activated protein5120176044247551764
kinase 10
ASJNK3(MAPK10)mitogen-activated protein5125646054229851765
kinase 10
ASJNK3(MAPK10)mitogen-activated protein5154006064241541766
kinase 10
ASMAP1mannan-binding lectin serine1692936071692931767
peptidase 1 (C4/C2 activating
component of Ra-reactive
factor)
ASMAP1mannan-binding lectin serine2962806082962801768
peptidase 1 (C4/C2 activating
component of Ra-reactive
factor)
ASMAP1mannan-binding lectin serine3377746093367921769
peptidase 1 (C4/C2 activating
component of Ra-reactive
factor)
ASMAP1mannan-binding lectin serine3924726103762641770
peptidase 1 (C4/C2 activating
component of Ra-reactive
factor)
ASMAP1mannan-binding lectin serine5418116114404461771
peptidase 1 (C4/C2 activating
component of Ra-reactive
factor)
ASMAP1mannan-binding lectin serine5418966124462401772
peptidase 1 (C4/C2 activating
component of Ra-reactive
factor)
ASMcl-1myeloid cell leukemia3079406133099731773
sequence 1 (BCL2-related)
ASMcl-1myeloid cell leukemia3690266143580221774
sequence 1 (BCL2-related)
ASMcl-1myeloid cell leukemia4397496154113951775
sequence 1 (BCL2-related)
ASMEK1mitogen-activated protein2158326162158321776
(MAP2K1)kinase 1
ASMEK1mitogen-activated protein3071026173024861777
(MAP2K1)kinase kinase 1
ASMEK1mitogen-activated protein4159116184091491778
(MAP2K1)kinase 1
ASMEK1mitogen-activated protein5447866194408421779
(MAP2K1)kinase 1
ASMEK2mitogen-activated protein2629486202629481780
(MAP2K2)kinase kinase 2
ASMEK4mitogen-activated protein3535336212624451781
(MAP2K4)kinase kinase 4
ASMEK4mitogen-activated protein4153856224104021782
(MAP2K4)kinase kinase 4
ASMEK4mitogen-activated protein5364136234416101783
(MAP2K4)kinase kinase 4
ASMEK4mitogen-activated protein5384656244448741784
(MAP2K4)kinase kinase 4
ASMEKK1mitogen-activated protein3995036253824231785
(MAP3K1)kinase kinase kinase 1
ASNADEnerve growth factor receptor2998726262998721786
(NGFRAP1)(TNFRSF16) associated
protein 1
ASNADEnerve growth factor receptor3612986273548431787
(NGFRAP1)(TNFRSF16) associated
protein 1
ASNADEnerve growth factor receptor3726346283617171788
(NGFRAP1)(TNFRSF16) associated
protein 1
ASNADEnerve growth factor receptor3726356293617181789
(NGFRAP1)(TNFRSF16) associated
protein 1
ASNADEnerve growth factor receptor3726456303617281790
(NGFRAP1)(TNFRSF16) associated
protein 1
ASNGFnerve growth factor (beta3695126313585251791
polypeptide)
ASNGFRnerve growth factor receptor1722296321722291792
ASNGFRnerve growth factor receptor5042016334217311793
ASNIKmitogen-activated protein3446866343420591794
(MAP3K14)kinase kinase kinase 14
ASNIKmitogen-activated protein3769266353661251795
(MAP3K14)kinase kinase kinase 14
ASNOXAphorbol-12-myristate-13-2695186362695181796
acetate-induced protein 1
ASNOXAphorbol-12-myristate-13-3166606373261191797
acetate-induced protein 1
ASOX40tumor necrosis factor receptor3792366383685381798
superfamily, member 4
ASOX40tumor necrosis factor receptor4535806393909071799
superfamily, member 4
ASOX40Ltumor necrosis factor (ligand)2818346402818341800
(TNFSF4)superfamily, member 4
ASOX40Ltumor necrosis factor (ligand)3677186413566911801
(TNFSF4)superfamily, member 4
ASOX40Ltumor necrosis factor (ligand)5452926424397041802
(TNFSF4)superfamily, member 4
ASp53tumor protein p532693056432693051803
ASp53tumor protein p5326930564426930518042489
ASp53tumor protein p533595976453526101805
ASp53tumor protein p533964736463797351806
ASp53tumor protein p534134656474107391807
ASp53tumor protein p534143156483941951808
ASp53tumor protein p534190246494021301809
ASp53tumor protein p534202466503911271810
ASp53tumor protein p5344588865139147818112490
ASp53tumor protein p534552636523988461812
ASp53tumor protein p535035916534262521813
ASp53tumor protein p535087936544241041814
ASp53tumor protein p535096906554251041815
ASp53tumor protein p535149446564238621816
ASp53tumor protein p535458586574377921817
ASp70 S6ribosomal protein S6 kinase,2255776582255771818
kinase 170 kDa, polypeptide 1
ASp70 S6ribosomal protein S6 kinase,3930216593767441819
kinase 170 kDa, polypeptide 1
ASp70 S6ribosomal protein S6 kinase,4061166603843351820
kinase 170 kDa, polypeptide 1
ASp70 S6ribosomal protein S6 kinase,4435726614419931821
kinase 170 kDa, polypeptide 1
ASp70 S6ribosomal protein S6 kinase,3126296623084131822
kinase 270 kDa, polypeptide 2
ASp70 S6ribosomal protein S6 kinase,5289646634328471823
kinase 270 kDa, polypeptide 2
ASp70 S6ribosomal protein S6 kinase,5391886644429491824
kinase 270 kDa, polypeptide 2
ASp90Rskribosomal protein S6 kinase,3741626653632771825
90 kDa, polypeptide 1
ASp90Rskribosomal protein S6 kinase,3741646663632791826
90 kDa, polypeptide 1
ASp90Rskribosomal protein S6 kinase,3741686673632831827
90 kDa, polypeptide 1
ASp90Rskribosomal protein S6 kinase,4037326683839671828
90 kDa, polypeptide 1
ASp90Rskribosomal protein S6 kinase,5300036694322811829
90 kDa, polypeptide 1
ASp90Rskribosomal protein S6 kinase,5313826704354121830
90 kDa, polypeptide 1
ASPAK2p21 protein (Cdc42/Rac)-3271346713140671831
activated kinase 2
ASPARP-1poly (ADP-ribose) polymerase 13667906723557551832
ASPARP-1poly (ADP-ribose) polymerase 13667916733557561833
ASPARP-1poly (ADP-ribose) polymerase 13667926743557571834
ASPARP-1poly (ADP-ribose) polymerase 13667946753557591835
ASPARP-1poly (ADP-ribose) polymerase 14323386764127741836
ASPDPK13-phosphoinositide dependent3420856773442201837
protein kinase-1
ASPDPK13-phosphoinositide dependent3548366783468951838
protein kinase-1
ASPDPK13-phosphoinositide dependent4415496793953571839
protein kinase-1
ASPI3Kphosphoinositide-3-kinase,2639676802639671840
catalytic, alpha polypeptide
ASPI3Kphosphoinositide-3-kinase,2891536812891531841
catalytic, beta polypeptide
ASPI3Kphosphoinositide-3-kinase,3591956823521211842
catalytic, gamma polypeptide
ASPI3Kphosphoinositide-3-kinase,3605636833537661843
catalytic, delta polypeptide
ASPI3Kphosphoinositide-3-kinase,3611106843544101844
catalytic, delta polypeptide
ASPI3Kphosphoinositide-3-kinase,3773466853665631845
catalytic, delta polypeptide
ASPI3Kphosphoinositide-3-kinase,4406506863922581846
catalytic, gamma polypeptide
ASPI3Kphosphoinositide-3-kinase,4614516874203991847
catalytic, beta polypeptide
ASPI3Kphosphoinositide-3-kinase,4680366884174791848
catalytic, alpha polypeptide
ASPI3Kphosphoinositide-3-kinase,4775936894181431849
catalytic, beta polypeptide
ASPI3Kphosphoinositide-3-kinase,4839686904198571850
catalytic, beta polypeptide
ASPI3Kphosphoinositide-3-kinase,4935686914178691851
catalytic, beta polypeptide
ASPI3Kphosphoinositide-3-kinase,4961666924192601852
catalytic, gamma polypeptide
ASPI3Kphosphoinositide-3-kinase,5366566934464441853
catalytic, delta polypeptide
ASPI3Kphosphoinositide-3-kinase,5433906944438111854
catalytic, delta polypeptide
ASPI3Kphosphoinositide-3-kinase,5447166954382591855
catalytic, beta polypeptide
ASPKA-catprotein kinase, cAMP-3086776963095911856
dependent, catalytic, alpha
ASPKA-catprotein kinase, cAMP-3503566973409401857
dependent, catalytic, alpha
ASPKA-catprotein kinase, cAMP-3706796983597131858
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-3706806993597141859
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-3706817003597151860
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-3706827013597161861
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-3706847023597181862
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-3706857033597191863
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-3706887043597221864
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-3706897053597231865
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-3772767063664881866
dependent, catalytic, gamma
ASPKA-catprotein kinase, cAMP-3948387073783141867
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-3948397083783151868
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-4135387093971751869
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-4175307103993261870
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-4321117113922751871
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-4361337123909061872
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-4465387134012521873
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-4507307143936541874
dependent, catalytic, beta
ASPKA-catprotein kinase, cAMP-5356957154416541875
dependent, catalytic, alpha
ASPKA-catprotein kinase, cAMP-5366497164404181876
dependent, catalytic, alpha
ASPKC-protein kinase C, delta3304527173316021877
delta
ASPKC-protein kinase C, delta3947297183782171878
delta
ASPKC-protein kinase C, delta4788437194197261879
delta
ASPKC-protein kinase C, delta4878977204181061880
delta
ASPKC-protein kinase C, zeta3785677213678301881
Zeta
ASPKC-protein kinase C, zeta4009207223837111882
Zeta
ASPKC-protein kinase C, zeta4009217233837121883
Zeta
ASPKC-protein kinase C, zeta4611067244264121884
Zeta
ASPKC-protein kinase C, zeta4705117254213501885
Zeta
ASPKC-protein kinase C, zeta4705967264242281886
Zeta
ASPKC-protein kinase C, zeta4709867274212191887
Zeta
ASPKC-protein kinase C, zeta4826867284253171888
Zeta
ASPKC-protein kinase C, zeta4963257294218691889
Zeta
ASPP1-catprotein phosphatase 1, catalytic3129897303260311890
alphasubunit, alpha isozyme
ASPP1-catprotein phosphatase 1, catalytic3767457313659361891
alphasubunit, alpha isozyme
ASPP1-catprotein phosphatase 1, catalytic4514587324056031892
alphasubunit, alpha isozyme
ASPP2aprotein phosphatase 2, catalytic4811957334184471893
catalyticsubunit, alpha isozyme
ASPP2Cprotein phosphatase,2287057342287051894
Mg2+/Mn2+ dependent, 1H
ASPP2Cprotein phosphatase,2632127352632121895
Mg2+/Mn2+ dependent, 1F
ASPP2Cprotein phosphatase,2824127362824121896
Mg2+/Mn2+ dependent, 1B
ASPP2Cprotein phosphatase,2959087372959081897
Mg2+/Mn2+ dependent, 1K
ASPP2Cprotein phosphatase,2964877382964871898
Mg2+/Mn2+ dependent, 1M
ASPP2Cprotein phosphatase,3059217393066821899
Mg2+/Mn2+ dependent, 1D
ASPP2Cprotein phosphatase,3082497403124111900
Mg2+/Mn2+ dependent, 1E
ASPP2Cprotein phosphatase,3092767413089261901
Mg2+/Mn2+ dependent, 1J
ASPP2Cprotein phosphatase,3151947423247611902
Mg2+/Mn2+ dependent, 1K
ASPP2Cprotein phosphatase,3235887433198941903
Mg2+/Mn2+ dependent, 1M
ASPP2Cprotein phosphatase,3246887443217611904
Mg2+/Mn2+ dependent, 1N
(putative)
ASPP2Cprotein phosphatase,3256427453272551905
Mg2+/Mn2+ dependent, 1A
ASPP2Cprotein phosphatase,3256587463148501906
Mg2+/Mn2+ dependent, 1A
ASPP2Cprotein phosphatase,3440347473427781907
Mg2+/Mn2+ dependent, 1G
ASPP2Cprotein phosphatase,3452497483260891908
Mg2+/Mn2+ dependent, 1B
ASPP2Cprotein phosphatase,3508037492647141909
Mg2+/Mn2+ dependent, 1G
ASPP2Cprotein phosphatase,3599947503530881910
Mg2+/Mn2+ dependent, 1J
ASPP2Cprotein phosphatase,3785517513678131911
Mg2+/Mn2+ dependent, 1B
ASPP2Cprotein phosphatase,3929957523767201912
Mg2+/Mn2+ dependent, 1D
ASPP2Cprotein phosphatase,3950767533785141913
Mg2+/Mn2+ dependent, 1A
ASPP2Cprotein phosphatase,3955437543789131914
Mg2+/Mn2+ dependent, 1G
ASPP2Cprotein phosphatase,3967347553799601915
Mg2+/Mn2+ dependent, 1N
(putative)
ASPP2Cprotein phosphatase,3974957563806321916
Mg2+/Mn2+ dependent, 1F
ASPP2Cprotein phosphatase,4069817573847151917
Mg2+/Mn2+ dependent, 1F
ASPP2Cprotein phosphatase,4071427583849301918
Mg2+/Mn2+ dependent, 1F
ASPP2Cprotein phosphatase,4094327593872871919
Mg2+/Mn2+ dependent, 1B
ASPP2Cprotein phosphatase,4095027603870461920
Mg2+/Mn2+ dependent, 1M
ASPP2Cprotein phosphatase,4098957613873411921
Mg2+/Mn2+ dependent, 1B
ASPP2Cprotein phosphatase,4198077623900871922
Mg2+/Mn2+ dependent, 1B
ASPP2Cprotein phosphatase,4431217633902571923
Mg2+/Mn2+ dependent, 1E
ASPP2Cprotein phosphatase,4573517643937471924
Mg2+/Mn2+ dependent, 1M
ASPP2Cprotein phosphatase,4973437654203541925
Mg2+/Mn2+ dependent, 1L
ASPP2Cprotein phosphatase,4981657664176591926
Mg2+/Mn2+ dependent, 1L
ASPP2Cprotein phosphatase,5064237674241551927
Mg2+/Mn2+ dependent, 1K
ASPP2Cprotein phosphatase,5253997684353981928
Mg2+/Mn2+ dependent, 1A
ASPP2Cprotein phosphatase,5282417694314531929
Mg2+/Mn2+ dependent, 1A
ASPP2Cprotein phosphatase,5295747704329661930
Mg2+/Mn2+ dependent, 1A
ASPP2Cprotein phosphatase,5319377714355751931
Mg2+/Mn2+ dependent, 1A
ASPP2Cprotein phosphatase,5381917724399151932
Mg2+/Mn2+ dependent, 1F
ASPP2Cprotein phosphatase,5444127734425361933
Mg2+/Mn2+ dependent, 1G
ASPP2Cprotein phosphatase,5447127744385181934
Mg2+/Mn2+ dependent, 1D
ASPumaBCL2 binding component 33008807753008801935
ASPumaBCL2 binding component 33419837763411551936
ASPumaBCL2 binding component 34390967773958621937
ASPumaBCL2 binding component 34492287784045031938
ASRAIDDCASP2 and RIPK1 domain3328967793276471939
containing adaptor with death
domain
ASRAIDDCASP2 and RIPK1 domain5418137804426241940
containing adaptor with death
domain
ASRAIDDCASP2 and RIPK1 domain5428937814390681941
containing adaptor with death
domain
ASRAIDDCASP2 and RIPK1 domain5510657824484251942
containing adaptor with death
domain
ASRANKtumor necrosis factor receptor2694857832694851943
superfamily, member 11a,
NFKB activator
ASRANKtumor necrosis factor receptor3827907843722401944
superfamily, member 11a,
NFKB activator
ASRANKLtumor necrosis factor (ligand)2398497852398491945
superfamily, member 11
ASRANKLtumor necrosis factor (ligand)3585457863513471946
superfamily, member 11
ASRANKLtumor necrosis factor (ligand)3987957873817751947
superfamily, member 11
ASRANKLtumor necrosis factor (ligand)4052627883840421948
superfamily, member 11
ASRANKLtumor necrosis factor (ligand)5448627894449131949
superfamily, member 11
ASReIAv-rel reticuloendotheliosis viral3086397903115081950
(p65oncogene homolog A (avian)
NF-
kappaB
subunit)
ASReIAv-rel reticuloendotheliosis viral4062467913842731951
(p65oncogene homolog A (avian)
NF-
kappaB
subunit)
ASReIAv-rel reticuloendotheliosis viral4266177924379801952
(p65oncogene homolog A (avian)
NF-
kappaB
subunit)
ASReIAv-rel reticuloendotheliosis viral5256937934325371953
(p65oncogene homolog A (avian)
NF-
kappaB
subunit)
ASReIAv-rel reticuloendotheliosis viral5262837944352901954
(p65oncogene homolog A (avian)
NF-
kappaB
subunit)
ASReIAv-rel reticuloendotheliosis viral5458167954437001955
(p65oncogene homolog A (avian)
NF-
kappaB
subunit)
ASRIPK1receptor (TNFRSF)-interacting2598087962598081956
serine-threonine kinase 1
ASRIPK1receptor (TNFRSF)-interacting3804097973697731957
serine-threonine kinase 1
ASRIPK1receptor (TNFRSF)-interacting4534837984159811958
serine-threonine kinase 1
ASRIPK1receptor (TNFRSF)-interacting5417917994422941959
serine-threonine kinase 1
ASSequestosome 1sequestosome 13607188003539441960
(p62)
ASSequestosome 1sequestosome 13769298013661281961
(p62)
ASSequestosome 1sequestosome 13898058023744551962
(p62)
ASSequestosome 1sequestosome 14028748033855531963
(p62)
ASSequestosome 1sequestosome 14222458043945341964
(p62)
ASSequestosome 1sequestosome 14543788054081071965
(p62)
ASSequestosome 1sequestosome 15140938064273081966
(p62)
ASShcSHC (Src homology 2 domain2645548072645541967
containing) transforming
protein 2
ASShcSHC (Src homology 2 domain3664428083961621968
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain3684418093574261969
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain3684438103574281970
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain3684458113574301971
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain3684498123574341972
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain3684508133574351973
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain3684538143574381974
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain3758308153649901975
containing) transforming
protein 3
ASShcSHC (Src homology 2 domain3758318163649911976
containing) transforming
protein 3
ASShcSHC (Src homology 2 domain3758358173649951977
containing) transforming
protein 3
ASShcSHC (Src homology 2 domain4121708183984411978
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain4141158194049081979
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain4441798203988641980
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain4446648213963331981
containing) transforming
protein 1
ASShcSHC (Src homology 2 domain4481168224013031982
containing) transforming
protein 1
ASSiah-1seven in absentia homolog 13567218233491561983
(Drosophila)
ASSiah-1seven in absentia homolog 13800068243693431984
(Drosophila)
ASSiah-1seven in absentia homolog 13947258253782141985
(Drosophila)
ASSMACdiablo, IAP-bindingNA826NA1986
mitochondrial protein
ASSmac/Diablodiablo, IAP-binding2671698272671691987
mitochondrial protein
ASSmac/Diablodiablo, IAP-binding3535488283203431988
mitochondrial protein
ASSmac/Diablodiablo, IAP-binding4139188294116381989
mitochondrial protein
ASSmac/Diablodiablo, IAP-binding4436498303984951990
mitochondrial protein
ASSmac/Diablodiablo, IAP-binding4649428314423601991
mitochondrial protein
ASSODDBCL2-associated athanogene 42873228322873221992
ASSODDBCL2-associated athanogene 44324718333932981993
ASSOSson of sevenless homolog 22163738342163731994
(Drosophila)
ASSOSson of sevenless homolog 12638798352638791995
(Drosophila)
ASSOSson of sevenless homolog 13950388363784791996
(Drosophila)
ASSOSson of sevenless homolog 14022198373846751997
(Drosophila)
ASSOSson of sevenless homolog 14260168383877841998
(Drosophila)
ASSOSson of sevenless homolog 14287218393999921999
(Drosophila)
ASSOSson of sevenless homolog 25436808404453282000
(Drosophila)
ASSOSson of sevenless homolog 15436988414411722001
(Drosophila)
ASSUMO-1SMT3 suppressor of mif two 33922448423760752002
homolog 1 (S. cerevisiae)
ASSUMO-1SMT3 suppressor of mif two 33922458433760762003
homolog 1 (S. cerevisiae)
ASSUMO-1SMT3 suppressor of mif two 33922468443760772004
homolog 1 (S. cerevisiae)
ASSUMO-1SMT3 suppressor of mif two 34092058453862672005
homolog 1 (S. cerevisiae)
ASSUMO-1SMT3 suppressor of mif two 34094988463864722006
homolog 1 (S. cerevisiae)
ASSurvivinbaculoviral IAP repeat3016338473016332007
containing 5
ASSurvivinbaculoviral IAP repeat3500518483241802008
containing 5
ASSurvivinbaculoviral IAP repeat3749488493640862009
containing 5
ASSurvivinbaculoviral IAP repeat4320148503890882010
containing 5
ASTACItumor necrosis factor receptor2616528512616522011
superfamily, member 13B
ASTACItumor necrosis factor receptor4375388524134532012
superfamily, member 13B
AStBidtBIDNANA2013
ASTL1Atumor necrosis factor (ligand)3740448533631562014
superfamily, member 15
ASTL1Atumor necrosis factor (ligand)3740458543631572015
superfamily, member 15
ASTNF-tumor necrosis factor3761228553652902016
alpha
ASTNF-tumor necrosis factor3834968563729882017
alpha
ASTNF-tumor necrosis factor4122758573928582018
alpha
ASTNF-tumor necrosis factor4204258584106682019
alpha
ASTNF-tumor necrosis factor4437078593894922020
alpha
ASTNF-tumor necrosis factor4452328603892652021
alpha
ASTNF-tumor necrosis factor4487818613894902022
alpha
ASTNF-tumor necrosis factor4492648623986982023
alpha
ASTNF-R1tumor necrosis factor receptor1627498631627492024
superfamily, member 1A
ASTNF-R1tumor necrosis factor receptor3661598643803892025
superfamily, member 1A
ASTNF-R2tumor necrosis factor receptor3762598653654352026
superfamily, member 1B
ASTNF-R2tumor necrosis factor receptor37625986636543520272491
superfamily, member 1B
ASTNF-R2tumor necrosis factor receptor4008638673836602028
superfamily, member 1B
ASTNF-R2tumor necrosis factor receptor5367828684404252029
superfamily, member 1B
ASTRADDTNFRSF1A-associated via3450578693412682030
death domain
ASTRAF2TNF receptor-associated factor 22476688702476682031
ASTRAF2TNF receptor-associated factor 23596628713526852032
ASTRAF2TNF receptor-associated factor 23716458723607082033
ASTRAF2TNF receptor-associated factor 24145898733976532034
ASTRAF2TNF receptor-associated factor 24190578744058602035
ASTRAF2TNF receptor-associated factor 24295098754065242036
ASTRAF2TNF receptor-associated factor 24327858764000612037
ASTRAF2TNF receptor-associated factor 25364688774464142038
ASTRAF3TNF receptor-associated factor 23476628783280032039
ASTRAF3TNF receptor-associated factor 33516918793324682040
ASTRAF3TNF receptor-associated factor 33927458803765002041
ASTRAF3TNF receptor-associated factor 35397218814459982042
ASTRAF3TNF receptor-associated factor 35603718824542072043
ASTRAF3TNF receptor-associated factor 35604638834536232044
ASTRAF5TNF receptor-associated factor 52614648842614642045
ASTRAF5TNF receptor-associated factor 53361848853368252046
ASTRAF5TNF receptor-associated factor 53670048863559712047
ASTRAF5TNF receptor-associated factor 54279258873898912048
ASTRAF6TNF receptor-associated factor 63481248883378532049
ASTRAF6TNF receptor-associated factor 65269958894336232050
ASTrkAneurotrophic tyrosine kinase,3681968903571792051
receptor, type 1
ASTrkAneurotrophic tyrosine kinase,3923028913761202052
receptor, type 1
ASTrkAneurotrophic tyrosine kinase,5243778924314182053
receptor, type 1
ASTWEAKtumor necrosis factor (ligand)2938258932938252054
(TNFSF12)superfamily, member 12
ASTWEAKtumor necrosis factor (ligand)5572338944514512055
(TNFSF12)superfamily, member 12
ASVDAC 1voltage-dependent anion2653338952653332056
channel 1
ASVDAC 1voltage-dependent anion3950448963784842057
channel 1
ASVDAC 1voltage-dependent anion3950478973784872058
channel 1
ASVDAC 2voltage-dependent anion2984688982984682059
channel 2
ASVDAC 2voltage-dependent anion3131328993616352060
channel 2
ASVDAC 2voltage-dependent anion3322119003616862061
channel 2
ASVDAC 2voltage-dependent anion3440369013448762062
channel 2
ASVDAC 2voltage-dependent anion4132899023895512063
channel 2
ASVDAC 2voltage-dependent anion4476779034014922064
channel 2
ASVDAC 2voltage-dependent anion5355539044459012065
channel 2
ASVDAC 2voltage-dependent anion5433519054430922066
channel 2
ASXIAPX-linked inhibitor of apoptosis3556409063478582067
ASXIAPX-linked inhibitor of apoptosis3711999073602422068
ASXIAPX-linked inhibitor of apoptosis4306259084006372069
ASXIAPX-linked inhibitor of apoptosis4347539093952302070
ASXIAPX-linked inhibitor of apoptosisNA910NA2071
CC/SATMataxia telangiectasia mutated2786169112786162072
CC/SATMataxia telangiectasia mutated3895119123741622073
CC/SATMataxia telangiectasia mutated4525089133880582074
CC/SATMataxia telangiectasia mutated5329319144323182075
CC/SATRataxia telangiectasia and Rad33507219153437412076
related
CC/SATRataxia telangiectasia and Rad33831019163725812077
related
CC/SATRIPATR interacting protein3202119173230992078
CC/SATRIPATR interacting protein3466919183023382079
CC/SATRIPATR interacting protein3571059193496202080
CC/SATRIPATR interacting protein4120529204009302081
CC/SATRIPATR interacting protein4211759214066642082
CC/SBard1BRCA1 associated RING2609479222609472083
domain 1
CC/SBard1BRCA1 associated RING4499679234067522084
domain 1
CC/SBLMBloom syndrome, RecQ3551129243472322085
helicase-like
CC/SBLMBloom syndrome, RecQ5369259254423302086
helicase-like
CC/SBLMBloom syndrome, RecQ5439779264390752087
helicase-like
CC/SBrca1breast cancer 1, early onset3094869273109382088
CC/SBrca1breast cancer 1, early onset3463159282469072089
CC/SBrca1breast cancer 1, early onset3516669293380072090
CC/SBrca1breast cancer 1, early onset3529939303122362091
CC/SBrca1breast cancer 1, early onset3540719313260022092
CC/SBrca1breast cancer 1, early onset3576549323502832093
CC/SBrca1breast cancer 1, early onset3936919333772942094
CC/SBrca1breast cancer 1, early onset4120619343971452095
CC/SBrca1breast cancer 1, early onset4612219354185482096
CC/SBrca1breast cancer 1, early onset4617989364179882097
CC/SBrca1breast cancer 1, early onset4683009374171482098
CC/SBrca1breast cancer 1, early onset4700269384192742099
CC/SBrca1breast cancer 1, early onset4711819394189602100
CC/SBrca1breast cancer 1, early onset4767779404175542101
CC/SBrca1breast cancer 1, early onset4771529414199882102
CC/SBrca1breast cancer 1, early onset4785319424204122103
CC/SBrca1breast cancer 1, early onset4840879434194812104
CC/SBrca1breast cancer 1, early onset4890379444207812105
CC/SBrca1breast cancer 1, early onset4917479454207052106
CC/SBrca1breast cancer 1, early onset4928599464202532107
CC/SBrca1breast cancer 1, early onset4937959474187752108
CC/SBrca1breast cancer 1, early onset4939199484188192109
CC/SBrca1breast cancer 1, early onset4941239494191032110
CC/SBrca1breast cancer 1, early onset4974889504189862111
CC/Sc-Ablc-abl oncogene 1, non-receptor3185609513233152112
tyrosine kinase
CC/Sc-Ablc-abl oncogene 1, non-receptor3723489523614232113
tyrosine kinase
CC/Sc-Ablc-abl oncogene 1, non-receptor3932939533769712114
tyrosine kinase
CC/Sc-Ablc-abl oncogene 1, non-receptor4384269544077562115
tyrosine kinase
CC/Sc-Ablc-abl oncogene 1, non-receptor4449709554004122116
tyrosine kinase
CC/SCDC25Acell division cycle 25 homolog3025069563037062117
A (S. pombe)
CC/SCDC25Acell division cycle 25 homolog3512319573431662118
A (S. pombe)
CC/SCDC25Acell division cycle 25 homolog4379729584042852119
A (S. pombe)
CC/SCDC25Bcell division cycle 25 homolog2459609592459602120
B (S. pombe)
CC/SCDC25Bcell division cycle 25 homolog3408339603391702121
B (S. pombe)
CC/SCDC25Bcell division cycle 25 homolog3442569613391252122
B (S. pombe)
CC/SCDC25Bcell division cycle 25 homolog3795989623689182123
B (S. pombe)
CC/SCDC25Bcell division cycle 25 homolog4398809634059722124
B (S. pombe)
CC/SCDC25Ccell division cycle 25 homolog3237609643216562125
C (S. pombe)
CC/SCDC25Ccell division cycle 25 homolog3489839653452052126
C (S. pombe)
CC/SCDC25Ccell division cycle 25 homolog3565059663488982127
C (S. pombe)
CC/SCDC25Ccell division cycle 25 homolog3572749673498212128
C (S. pombe)
CC/SCDC25Ccell division cycle 25 homolog4151309683926312129
C (S. pombe)
CC/SCDC25Ccell division cycle 25 homolog5030229694272512130
C (S. pombe)
CC/SCDC25Ccell division cycle 25 homolog5139709704247952131
C (S. pombe)
CC/SCDC25Ccell division cycle 25 homolog5348929714431962132
C (S. pombe)
CC/SCDK2cyclin-dependent kinase 22669709722669702133
CC/SCDK2cyclin-dependent kinase 23540569732430672134
CC/SCDK4cyclin-dependent kinase 42579049742579042135
CC/SCDK4cyclin-dependent kinase 43129909753168892136
CC/SCDK4cyclin-dependent kinase 45403259764390762137
CC/SCDK4cyclin-dependent kinase 45522549774491792138
CC/SCDK4cyclin-dependent kinase 45523889784489632139
CC/SCDK4cyclin-dependent kinase 45528629794467632140
CC/SCDK6cyclin-dependent kinase 62657349802657342141
CC/SCDK6cyclin-dependent kinase 64248489813970872142
CC/SChk1checkpoint kinase 12789169822789162143
CC/SChk1checkpoint kinase 14288309834125042144
CC/SChk1checkpoint kinase 14380159843886482145
CC/SChk1checkpoint kinase 15247379854328902146
CC/SChk1checkpoint kinase 15253969864341412147
CC/SChk1checkpoint kinase 15269379874318152148
CC/SChk1checkpoint kinase 15270139884315252149
CC/SChk1checkpoint kinase 15340709894353712150
CC/SChk1checkpoint kinase 15346859904324702151
CC/SChk1checkpoint kinase 15443739914423172152
CC/SChk2checkpoint kinase 23283549923291782153
CC/SChk2checkpoint kinase 23482959933290122154
CC/SChk2checkpoint kinase 23825639943720032155
CC/SChk2checkpoint kinase 23825659953720062156
CC/SChk2checkpoint kinase 23825669963720072157
CC/SChk2checkpoint kinase 23825789973720212158
CC/SChk2checkpoint kinase 23825809983720232159
CC/SChk2checkpoint kinase 24027319993848352160
CC/SChk2checkpoint kinase 240364210003849192161
CC/SChk2checkpoint kinase 240427610013857472162
CC/SChk2checkpoint kinase 240559810023860872163
CC/SChk2checkpoint kinase 254477210034424582164
CC/SClaspinclaspin25119510042511952165
CC/SClaspinclaspin31812110053129952166
CC/SClaspinclaspin37322010063623172167
CC/SClaspinclaspin54435610074423352168
CC/SCyclin Acyclin A227402610082740262169
CC/SCyclin Bcyclin B125644210092564422170
CC/SCyclin Bcyclin B327601410102760142171
CC/SCyclin Bcyclin B228820710112882072172
CC/SCyclin Bcyclin B334860310123386822173
CC/SCyclin Bcyclin B337603810133652062174
CC/SCyclin Bcyclin B337604210143652102175
CC/SCyclin Bcyclin B339654010153797902176
CC/SCyclin Bcyclin B150550010164245882177
CC/SCyclin Bcyclin B150657210174233872178
CC/SCyclin Dcyclin D122750710182275072179
CC/SCyclin Dcyclin D226125410192612542180
CC/SCyclin Dcyclin D337298710203620782181
CC/SCyclin Dcyclin D337298810213620792182
CC/SCyclin Dcyclin D337299110223620822183
CC/SCyclin Dcyclin D341420010233975452184
CC/SCyclin Dcyclin D341549710244015952185
CC/SCyclin Dcyclin D350506410254258302186
CC/SCyclin Dcyclin D351164210264262122187
CC/SCyclin Dcyclin D154289710274418632188
CC/SCyclin Ecyclin E126264310282626432189
CC/SCyclin Ecyclin E230810810293091812190
CC/SCyclin Ecyclin E135794310303506252191
CC/SCyclin Ecyclin E239613310313794372192
CC/SCyclin Ecyclin E144498310324101792193
CC/SCyclin Ecyclin E252050910334290892194
CC/SCyclin Ecyclin E254272510344457262195
CC/SDNA-protein kinase, DNA-activated,31419110353134202196
PKcatalytic polypeptide
CC/SDNA-protein kinase, DNA-activated,33836810363451822197
PKcatalytic polypeptide
CC/SE2F1/2/E2F transcription factor 5,25611710372561172198
3/4/5/6p130-binding
CC/SE2F1/2/E2F transcription factor 630723610383021592199
3/4/5/6
CC/SE2F1/2/E2F transcription factor 134338010393455712200
3/4/5/6
CC/SE2F1/2/E2F transcription factor 334661810402629042201
3/4/5/6
CC/SE2F1/2/E2F transcription factor 236172910413552492202
3/4/5/6
CC/SE2F1/2/E2F transcription factor 636200910423550362203
3/4/5/6
CC/SE2F1/2/E2F transcription factor 337864610433679142204
3/4/5/6
CC/SE2F1/2/E2F transcription factor 4,37937810443686862205
3/4/5/6p107/p130-binding
CC/SE2F1/2/E2F transcription factor 638152510453709362206
3/4/5/6
CC/SE2F1/2/E2F transcription factor 5,41627410463981242207
3/4/5/6p130-binding
CC/SE2F1/2/E2F transcription factor 5,41893010474143122208
3/4/5/6p130-binding
CC/SE2F1/2/E2F transcription factor 5,51747610484291202209
3/4/5/6p130-binding
CC/SE2F1/2/E2F transcription factor 5,51823410494296692210
3/4/5/6p130-binding
CC/SE2F1/2/E2F transcription factor 353543210504434182211
3/4/5/6
CC/SE2F1/2/E2F transcription factor 654210010514463152212
3/4/5/6
CC/SE2F1/2/E2F transcription factor 654621210524388642213
3/4/5/6
CC/SFANCD2Fanconi anemia,28764710532876472214
complementation group D2
CC/SFANCD2Fanconi anemia,38380610543733172215
complementation group D2
CC/SFANCD2Fanconi anemia,38380710553733182216
complementation group D2
CC/SFANCD2Fanconi anemia,41958510563987542217
complementation group D2
CC/SFANCLFanconi anemia,23374110572337412218
complementation group L
CC/SFANCLFanconi anemia,54064610584414312219
complementation group L
CC/SGADD4growth arrest and DNA-37098610593600252220
5 alphadamage-inducible, alpha
CC/SGADD4growth arrest and DNA-21563110602156312221
5 betadamage-inducible, beta
CC/SGADD4growth arrest and DNA-37098510613600242222
5 betadamage-inducible, alpha
CC/SMDM2Mdm2 p53 binding protein25814810622581482223
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein25814910632581492224
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein29925210642992522225
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein31142010653107422226
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein31144010663113022227
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein34880110673350962228
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein35005710682666242229
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein35629010693486372230
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein35848310703512702231
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein36043010713536112232
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein39341010723770622233
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein39341210733770642234
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein39341310743770652235
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein39341510753770672236
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein42886310764106942237
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein46228410774172812238
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein51785210784302572239
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein53947910794444302240
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein54082710804409322241
homolog (mouse)
CC/SMDM2Mdm2 p53 binding protein54464810814432742242
homolog (mouse)
CC/SNFBD1mediator of DNA-damage37640510823655872243
checkpoint 1
CC/SNFBD1mediator of DNA-damage37640610833655882244
checkpoint 1
CC/SNFBD1mediator of DNA-damage38356610843730602245
checkpoint 1
CC/SNFBD1mediator of DNA-damage41239510853928332246
checkpoint 1
CC/SNFBD1mediator of DNA-damage41397310864088312247
checkpoint 1
CC/SNFBD1mediator of DNA-damage41636810874103832248
checkpoint 1
CC/SNFBD1mediator of DNA-damage41657110884009792249
checkpoint 1
CC/SNFBD1mediator of DNA-damage41703310894089622250
checkpoint 1
CC/SNFBD1mediator of DNA-damage41722810904003052251
checkpoint 1
CC/SNFBD1mediator of DNA-damage41917210913984742252
checkpoint 1
CC/SNFBD1mediator of DNA-damage41967510923976422253
checkpoint 1
CC/SNFBD1mediator of DNA-damage42001910933964842254
checkpoint 1
CC/SNFBD1mediator of DNA-damage42032010944165112255
checkpoint 1
CC/SNFBD1mediator of DNA-damage42210410953903752256
checkpoint 1
CC/SNFBD1mediator of DNA-damage42219510964077032257
checkpoint 1
CC/SNFBD1mediator of DNA-damage42226610974113102258
checkpoint 1
CC/SNFBD1mediator of DNA-damage42372610983912302259
checkpoint 1
CC/SNFBD1mediator of DNA-damage42443710993981512260
checkpoint 1
CC/SNFBD1mediator of DNA-damage42450711003883552261
checkpoint 1
CC/SNFBD1mediator of DNA-damage42463811013940742262
checkpoint 1
CC/SNFBD1mediator of DNA-damage42502911023971262263
checkpoint 1
CC/SNFBD1mediator of DNA-damage42507211033969892264
checkpoint 1
CC/SNFBD1mediator of DNA-damage42579011043970212265
checkpoint 1
CC/SNFBD1mediator of DNA-damage42740611053874292266
checkpoint 1
CC/SNFBD1mediator of DNA-damage42961011064068502267
checkpoint 1
CC/SNFBD1mediator of DNA-damage43035811074141632268
checkpoint 1
CC/SNFBD1mediator of DNA-damage43144111083927842269
checkpoint 1
CC/SNFBD1mediator of DNA-damage43299811094059912270
checkpoint 1
CC/SNFBD1mediator of DNA-damage43566411104043182271
checkpoint 1
CC/SNFBD1mediator of DNA-damage43579711114006772272
checkpoint 1
CC/SNFBD1mediator of DNA-damage43775911123877432273
checkpoint 1
CC/SNFBD1mediator of DNA-damage43816511133877062274
checkpoint 1
CC/SNFBD1mediator of DNA-damage44036911144152122275
checkpoint 1
CC/SNFBD1mediator of DNA-damage44139711153904892276
checkpoint 1
CC/SNFBD1mediator of DNA-damage44441211164136102277
checkpoint 1
CC/SNFBD1mediator of DNA-damage44513011173961242278
checkpoint 1
CC/SNFBD1mediator of DNA-damage44576411183938862279
checkpoint 1
CC/SNFBD1mediator of DNA-damage44719211194058062280
checkpoint 1
CC/SNFBD1mediator of DNA-damage44764011203963892281
checkpoint 1
CC/SNFBD1mediator of DNA-damage44889511213961212282
checkpoint 1
CC/SNFBD1mediator of DNA-damage44915311224091672283
checkpoint 1
CC/SNFBD1mediator of DNA-damage45003311233900402284
checkpoint 1
CC/SNFBD1mediator of DNA-damage45221311244049362285
checkpoint 1
CC/SNFBD1mediator of DNA-damage45572911254049542286
checkpoint 1
CC/SNFBD1mediator of DNA-damage45658911264053502287
checkpoint 1
CC/SNFBD1mediator of DNA-damage54648711274486792288
checkpoint 1
CC/SNFBD1mediator of DNA-damage54653911284482322289
checkpoint 1
CC/SNFBD1mediator of DNA-damage54704711294490592290
checkpoint 1
CC/SNFBD1mediator of DNA-damage54735311304478832291
checkpoint 1
CC/SNFBD1mediator of DNA-damage54768111314478512292
checkpoint 1
CC/SNFBD1mediator of DNA-damage54770011324490832293
checkpoint 1
CC/SNFBD1mediator of DNA-damage54787411334476822294
checkpoint 1
CC/SNFBD1mediator of DNA-damage54810311344494992295
checkpoint 1
CC/SNFBD1mediator of DNA-damage54811211354484342296
checkpoint 1
CC/SNFBD1mediator of DNA-damage54824811364480802297
checkpoint 1
CC/SNFBD1mediator of DNA-damage54843311374499712298
checkpoint 1
CC/SNFBD1mediator of DNA-damage54854211384465972299
checkpoint 1
CC/SNFBD1mediator of DNA-damage54880511394469242300
checkpoint 1
CC/SNFBD1mediator of DNA-damage54882711404492012301
checkpoint 1
CC/SNFBD1mediator of DNA-damage54889311414479432302
checkpoint 1
CC/SNFBD1mediator of DNA-damage54894711424477112303
checkpoint 1
CC/SNFBD1mediator of DNA-damage54922811434475172304
checkpoint 1
CC/SNFBD1mediator of DNA-damage54938211444491772305
checkpoint 1
CC/SNFBD1mediator of DNA-damage54942811454470382306
checkpoint 1
CC/SNFBD1mediator of DNA-damage54977111464488122307
checkpoint 1
CC/SNFBD1mediator of DNA-damage55000411474470842308
checkpoint 1
CC/SNFBD1mediator of DNA-damage55011011484469802309
checkpoint 1
CC/SNFBD1mediator of DNA-damage55021011494476972310
checkpoint 1
CC/SNFBD1mediator of DNA-damage55040811504471362311
checkpoint 1
CC/SNFBD1mediator of DNA-damage55050011514500022312
checkpoint 1
CC/SNFBD1mediator of DNA-damage55068811524480662313
checkpoint 1
CC/SNFBD1mediator of DNA-damage55120411534477992314
checkpoint 1
CC/SNFBD1mediator of DNA-damage55126711544501982315
checkpoint 1
CC/SNFBD1mediator of DNA-damage55146011554492742316
checkpoint 1
CC/SNFBD1mediator of DNA-damage55155411564485382317
checkpoint 1
CC/SNFBD1mediator of DNA-damage55162111574482852318
checkpoint 1
CC/SNFBD1mediator of DNA-damage55174011584500372319
checkpoint 1
CC/SNFBD1mediator of DNA-damage55226311594470692320
checkpoint 1
CC/SNFBD1mediator of DNA-damage55234911604498922321
checkpoint 1
CC/SNFBD1mediator of DNA-damage55247411614477712322
checkpoint 1
CC/SNFBD1mediator of DNA-damage55252211624499362323
checkpoint 1
CC/SNFBD1mediator of DNA-damage55277611634478252324
checkpoint 1
CC/SNFBD1mediator of DNA-damage55304711644472472325
checkpoint 1
CC/SNFBD1mediator of DNA-damage55304811654477872326
checkpoint 1
CC/SNFBD1mediator of DNA-damage55313011664468092327
checkpoint 1
CC/SNFBD1mediator of DNA-damage55319611674495862328
checkpoint 1
CC/SNibrinnibrin26543311682654332329
CC/SNibrinnibrin45238711694452132330
CC/Sp107retinoblastoma-like 1 (p107)34435911703436462331
CC/Sp107retinoblastoma-like 1 (p107)37366411713627682332
CC/Sp130retinoblastoma-like 2 (p130)26213311722621332333
CC/Sp130retinoblastoma-like 2 (p130)37993511733692672334
CC/Sp130retinoblastoma-like 2 (p130)54440511744437442335
CC/Sp130retinoblastoma-like 2 (p130)54454511754446852336
CC/Sp21P21NA1176NA2337
CC/SPCNAproliferating cell nuclear37914311773684382338
antigen
CC/SPCNAproliferating cell nuclear37916011783684582339
antigen
CC/SRAD9RAD9 homolog A (S. pombe)30798011793113602340
CC/SRbretinoblastoma 126716311802671632341
protein
CC/SRbretinoblastoma 146750511814347022342
protein
CC/SRbretinoblastoma 154291711824376422343
protein
CC/SSMC1structural maintenance of32221311833234212344
chromosomes 1A
CC/SSMC1structural maintenance of34021311843449062345
chromosomes 1A
CC/SSMC1structural maintenance of37534011853644892346
chromosomes 1A
CC/SSMC1structural maintenance of42801411864135092347
chromosomes 1A
CC/SUSP1ubiquitin specific peptidase 133995011873435262348
CC/SUSP1ubiquitin specific peptidase 137114611883601882349
CC/SUSP1ubiquitin specific peptidase 145214311894036622350
M4EBP-1eukaryotic translation initiation33882511903406912351
factor 4E binding protein 1
MARNTaryl hydrocarbon receptor35439611913463722352
nuclear translocator
MARNTaryl hydrocarbon receptor35859511923514072353
nuclear translocator
MARNTaryl hydrocarbon receptor36897511933579712354
nuclear translocator
MARNTaryl hydrocarbon receptor39470011943781902355
nuclear translocator
MARNTaryl hydrocarbon receptor47184411954258992356
nuclear translocator
MARNTaryl hydrocarbon receptor50575511964275712357
nuclear translocator
MARNTaryl hydrocarbon receptor51519211974238512358
nuclear translocator
MCAIXcarbonic anhydrase IX37835711983676082359
MCAIXcarbonic anhydrase IX54407411994385412360
MCBPCREB binding protein26236712002623672361
MCBPCREB binding protein32350812013235502362
MCBPCREB binding protein38207012023715022363
MCBPCREB binding protein54388312034419782364
MCITED1Cbp/p300-interacting24613912042461392365
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 1
MCITED1Cbp/p300-interacting37361912053627212366
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 1
MCITED1Cbp/p300-interacting41740012064147812367
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 1
MCITED1Cbp/p300-interacting42741212073914072368
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 1
MCITED1Cbp/p300-interacting42979412084074962369
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 1
MCITED1Cbp/p300-interacting43138112093885482370
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 1
MCITED1Cbp/p300-interacting44598312104032742371
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 1
MCITED1Cbp/p300-interacting45087512114057652372
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 1
MCITED1Cbp/p300-interacting45370712124017642373
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 1
MCITED2Cbp/p300-interacting36765112133566232374
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 2
MCITED2Cbp/p300-interacting39231212143761262375
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 2
MCITED2Cbp/p300-interacting53615912154428312376
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 2
MCITED2Cbp/p300-interacting53733212164441982377
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 2
MCITED4Cbp/p300-interactingNA1217NA2378
transactivator, with Glu/Asp-
rich carboxy-terminal domain, 4
MCITED4Cbp/p300-interacting37263812183617212379
transactivator, with Glu/Asp-
rich carboxy-terminal domain,
4 (CBP/p300 interacting
transactivator with ED-rich
tail)
MCOMMD1copper metabolism (Murr1)31183212193082362380
domain containing 1
MCOMMD1copper metabolism (Murr1)42741712204132072381
domain containing 1
MCOMMD1copper metabolism (Murr1)44416612214100502382
domain containing 1
MCOMMD1copper metabolism (Murr1)45833712224012362383
domain containing 1
MCOMMD1copper metabolism (Murr1)53873612234389612384
domain containing 1
MCREBcAMP responsive element23699612242369962385
binding protein 1
MCREBcAMP responsive element35326712252369952386
binding protein 1
MCREBcAMP responsive element35370412263421362387
binding protein 3
MCREBcAMP responsive element37439712273635182388
binding protein 1
MCREBcAMP responsive element43062412284055392389
binding protein 1
MCREBcAMP responsive element43232912293876992390
binding protein 1
MCREBcAMP responsive element44580312304072272391
binding protein 1
MCREBcAMP responsive element45247412313924282392
binding protein 1
MCREBcAMP responsive element53672612324458922393
binding protein 1
MCREBcAMP responsive element53978912334408092394
binding protein 1
MeIF4Eeukaryotic translation initiation28089212342808922395
factor 4E
MeIF4Eeukaryotic translation initiation45025312353896242396
factor 4E
MHIF3-hypoxia inducible factor 3,24430212362443022397
alphaalpha subunit
MHIF3-hypoxia inducible factor 3,29130012372913002398
alphaalpha subunit
MFIHhypoxia inducible factor 1,29916312382991632399
alpha subunit inhibitor (factor
inhibiting HIF)
MHIF3-hypoxia inducible factor 3,30086212393008622400
alpha)alpha subunit
MHIF3-hypoxia inducible factor 3,33961312403418772401
alphaalpha subunit
MHIF3-hypoxia inducible factor 3,37767012413668982402
alphaalpha subunit
MHIF3-hypoxia inducible factor 3,41470712424128082403
alphaalpha subunit
MHIF3-hypoxia inducible factor 3,42010212434077712404
alphaalpha subunit
MFIHhypoxia inducible factor 1,44272412443997342405
(factoralpha subunit inhibitor
inhibiting
HIF)
MHIF3-hypoxia inducible factor 3,45777112454080082406
alphaalpha subunit
MHIF3-hypoxia inducible factor 3,45786512463940522407
alphaalpha subunit
MHIF3-hypoxia inducible factor 3,47543212474325782408
alphaalpha subunit
MFIHhypoxia inducible factor 1,53358912484333602409
(factoralpha subunit inhibitor
inhibiting
HIF)
MGrb2growth factor receptor-bound31661512493173602410
protein 2
MGrb2growth factor receptor-bound31680412503390072411
protein 2
MGrb2growth factor receptor-bound39256212513763452412
protein 2
MGrb2growth factor receptor-bound39256412523763472413
protein 2
MHNF4alphahepatocyte nuclear factor 4,31609912533129872414
alpha
MHNF4alphahepatocyte nuclear factor 4,31667312543151802415
alpha
MHNF4alphahepatocyte nuclear factor 4,33869212553438072416
alpha
MHNF4alphahepatocyte nuclear factor 4,41569112564121112417
alpha
MHNF4alphahepatocyte nuclear factor 4,44359812574109112418
alpha
MHNF4alphahepatocyte nuclear factor 4,45723212583962162419
alpha
MHNF4alpha2Homo sapiens hepatocyteNA1259NA2420
nuclear factor 4, alpha
(HNF4A), transcript variant 2,
mRNA
MIBP3insulin-like growth factor27552112602755212421
binding protein 3
MIBP3insulin-like growth factor38108312613704732422
binding protein 3
MIBP3insulin-like growth factor38108612623704762423
binding protein 3
MIBP3insulin-like growth factor41762112633991162424
binding protein 3
MIBP3insulin-like growth factor42853012643902982425
binding protein 3
MIBP3insulin-like growth factor43304712654044612426
binding protein 3
MIBP3insulin-like growth factor43849112663937402427
binding protein 3
MIBP3insulin-like growth factor44214212673924722428
binding protein 3
MIBP3insulin-like growth factor54503212684399992429
binding protein 3
MJAB1COP9 constitutive35784912693505122430
photomorphogenic homolog
subunit 5 (Arabidopsis)
MMNK1MAP kinase interacting34118312703395732431
serine/threonine kinase 1
MMNK1MAP kinase interacting37194412713610122432
serine/threonine kinase 1
MMNK1MAP kinase interacting37194512723610132433
serine/threonine kinase 1
MMNK1MAP kinase interacting37194612733610142434
serine/threonine kinase 1
MMNK1MAP kinase interacting42811212744111352435
serine/threonine kinase 1
MMNK1MAP kinase interacting49661912754367092436
serine/threonine kinase 1
MMNK1MAP kinase interacting54573012764409742437
serine/threonine kinase 1
MMNK2MAP kinase interacting25089612772508962438
serine/threonine kinase 2
MMNK2MAP kinase interacting30934012783094852439
serine/threonine kinase 2
MMNK2MAP kinase interacting54116512794389042440
serine/threonine kinase 2
MMNK2MAP kinase interacting54562712804412452441
serine/threonine kinase 2
Mp15(INK4A)cyclin-dependent kinase27692512812769252442
inhibitor 2B (p15, inhibits
CDK4)
Mp15(INK4A)cyclin-dependent kinase38014212823694872443
inhibitor 2B (p15, inhibits
CDK4)
Mp300E1A binding protein p30026325312832632532444
MPer1period homolog 1 (Drosophila)31727612843144202445
MPer1period homolog 1 (Drosophila)35490312853469792446
MRPS6ribosomal protein S631537712863697432447
MRPS6ribosomal protein S638038112873697412448
MRPS6ribosomal protein S638038412883697452449
MRPS6ribosomal protein S638039412893697572450
MSHARP1basic helix-loop-helix family,NA1290NA2451
member e41
MSHARP1basic helix-loop-helix family,24272812912427282452
(BHLHE41)member e41
MSHARP1basic helix-loop-helix family,54073112924373692453
(BHLHE41)member e41
MSRC1nuclear receptor coactivator 128859912932885992454
MSRC1nuclear receptor coactivator 134833212943209402455
MSRC1nuclear receptor coactivator 139585612953791972456
MSRC1nuclear receptor coactivator 140514112963850972457
MSRC1nuclear receptor coactivator 140696112973852162458
MSRC1nuclear receptor coactivator 153853912984440392459
Mtuberintuberous sclerosis 221947612992194762460
Mtuberintuberous sclerosis 235077313003443832461
Mtuberintuberous sclerosis 235392913012480992462
Mtuberintuberous sclerosis 238253813023719782463
Mtuberintuberous sclerosis 240187413033844682464
Mtuberintuberous sclerosis 243967313043992322465
AIFSHapoptosis-inducing factor,NA1305NA2466
short
Angiopoietin1Angiopoietin 1NA1306NA24672492
BMP2BMP2 CONA1307NA24682493
CO
c-MYCv-myc myelocytomatosis viralNA1308NA2469
oncogene homolog (avian)
COMMD1COMMD1NA1309NA
COMMD1COMMD1 with nuclear exportNANA2470
NESseqences deleted
deleted
COMMD1COMMD1 with nuclear exportNANA2471
NES1sequences deleted and nuclear
deletedlocalization signals added
and NLS
added
COMMD1COMMD1 with SV40 andNANA2472
SV40nuclear localization signals
NLS
COMMD1wtCOMMD1 wild-typeNANA2473
GLUT1solute carrier family 2NA1310NA2474
(facilitated glucose
transporter), member 1
GranulysinGranulysin FL15NA1311NA2475
FL15
GranulysinGranulysin NS9NANA24762494
NS9
GranulysinGranulysin S9NANA24772495
S9
HIF1 ahypoxia inducible factor 1,NA1312NA2478
alpha subunit (basic helix-
loop-helix transcription factor)
IL15interleukin 15NA1313NA2479
KGFfibroblast growth factor 7,NA1314NA2480
precursor; mature is 32-194
MCT4solute carrier family 16,NA1315NA24812496
member 4 (monocarboxylic
acid transporter 5)
MYCMYC inhibitor D (OMOMyc)NA1316NA2482
inhibitor D
MYCMYC inhibitor D_90NANA2483
inhibitor(OmoMyc_90)
D_90
C.A.Constitutively active (C.A.)NA1317NA2484
caspasecaspase 3 cleavable
3_cleavable(RevCasp3_Cleavable)
C.A.Constitutively active (C.A.)NA1318NA2485
caspasecaspase 3 uncleavable
3_uncleavable(RevCasp3_UnCleavable)
C.A.Constitutively active (C.A.)NA1319NA2486
caspase 6caspase 6 (RevCasp6)
SIAh1siah E3 ubiquitin protein ligase 1NA1320NA2487
HSV1-tkHerpes simplex virus 1-
thymidine kinase

Shown in Table 7, are familiar cancer syndromes, tumor suppressor genes, function of the tumor suppressor gene, chromosomal location, and tumor type observed. Signal-sensor polynucleotides of the present invention can be designed as a therapeutic for any of those listed in the table.

TABLE 7
Familial Cancer Syndrome Targets
FamilialTumor
CancerSuppressorChromosomalTumor Types
SyndromeGeneFunctionLocationObserved
Li-FraumeniP53cell cycle17p13.1brain tumors,
Syndromeregulation,sarcomas, leukemia,
apoptosisbreast cancer
FamilialRB1cell cycle13q14.1-q14.2retinoblastoma,
Retinoblastomaregulationosteogenic sarcoma
Wilms TumorWT1transcriptional11p13pediatric kidney
regulationcancer, most
common form of
childhood solid
tumor
NeurofibromatosisNF1catalysis of RAS17q11.2neurofibromas,
Type 1inactivationsarcomas, gliomas
NeurofibromatosisNF2linkage of cell22q12.2Schwann cell
Type 2membrane to actintumors,
cytoskeletonastrocytomas,
meningiomas,
ependymonas
FamilialAPCsignaling through5q21-q22colon cancer
Adenomatousadhesion
Polyposismolecules to
nucleus
TuberousTSC1forms complex9q34seizures, mental
sclerosis 1with TSC2retardation, facial
protein, inhibitsangiofibromas
signaling to
downstream
effectors of mTOR
TuberousTSC2forms complex16p13.3benign growths
sclerosis 2with TSC1(hamartomas) in
protein, inhibitsmany tissues,
signaling toastrocytomas,
downstreamrhabdomyosarcomas
effectors of mTOR
Deleted inDPC4, alsoregulation of18q21.1pancreatic
Pancreaticknown asTGF-β/BMPcarcinoma, colon
Carcinoma 4,SMAD4signal transductioncancer
Familial
juvenile
polyposis
syndrome
Deleted inDCCtransmembrane18q21.3colorectal cancer
Colorectalreceptor involved
Carcinomain axonal guidance
via netrins
Familial BreastBRCA1functions in17q21breast and ovarian
Cancertranscription,cancer
DNA binding,
transcription
coupled DNA
repair,
homologous
recombination,
chromosomal
stability,
ubiquitination of
proteins, and
centrosome
replication
Familial BreastBRCA2transcriptional13q12.3breast and ovarian
Cancer(FANCD1)regulation ofcancer
genes involved in
DNA repair and
homologous
recombination
CowdenPTENphosphoinositide10q23.3gliomas, breast
syndrome3-phosphatase,cancer, thyroid
protein tyrosinecancer, head & neck
phosphatasesquamous carcinoma
Peutz-JeghersSTK11phosphorylates19p13.3hyperpigmentation,
Syndrome (PJS)(serine-and activatesmultiple
threonineAMP-activatedhamartomatous
kinase 11)kinase (AMPK),polyps, colorectal,
AMPK involvedbreast and ovarian
in stresscancers
responses, lipid
and glucose
meatabolism
HereditaryMSH2DNA mismatch2p22-p21colon cancer
Nonpolyposisrepair
Colon Cancer
type 1,
HNPCC1
HereditaryMLH1DNA mismatch3p21.3colon cancer
Nonpolyposisrepair
Colon Cancer
type 2,
HNPCC2
Familial diffuse-CDH1cell-cell adhesion16q22.1gastric cancer,
type gastricproteinlobular breast cancer
cancer
von Hippel-VHLregulation of3p26-p25renal cancers,
Lindautranscriptionhemangioblastomas,
Syndromeelongation throughpheochromocytoma,
activation of aretinal angioma
ubiquitin ligase
complex
FamilialCDKN2Ap16INK49p21melanoma,
Melanomainhibits cell-cyclepancreatic cancer,
kinases CDK4 andothers
CDK6; p14ARF
binds
the p53 stabilizing
protein MDM2
GorlinPTCHtransmembrane9q22.3basal cell skin
Syndrome:(e.g.,receptor for soniccarcinoma
Nevoid basalPTCH1,hedgehog (shh),
cell carcinomaPTCH2)involved in early
syndromedevelopment
(NBCCS)through repression
of action of
smoothened
MultipleMEN1intrastrand DNA11q13parathyroid and
Endocrinecrosslink repairpituitary adenomas,
Neoplasia Typeislet cell tumors,
1carcinoid

In additional to the above mentioned targets, the oncology-related polypeptides may include any “death signal” protein that can be recognized by active T cells of immune system. Such suicide signal proteins encoded by the sensor-signal polynucleotides can be selectively expressed in particular tissues or cells (e.g. cancer cells) through engineered microRNA binding sites and/or other regulatory elements as described herein. The group of proteins, when they are expressed on the surface of a cancer cell, can prime T cell to induce T cell mediated immune response, thus killing the cancer cell. As a non-limiting example, a group of proteins that are known to present a “death signal”, include, CD80, CD86, B7 and MHC II, etc.

Protein Cleavage Signals and Sites

In one embodiment, the oncology-related polypeptides of the present invention may include at least one protein cleavage signal containing at least one protein cleavage site. The protein cleavage site may be located at the N-terminus, the C-terminus, at any space between the N- and the C-termini such as, but not limited to, half-way between the N- and C-termini, between the N-terminus and the half way point, between the half way point and the C-terminus, and combinations thereof.

The oncology-related polypeptides of the present invention may include, but is not limited to, a proprotein convertase (or prohormone convertase), thrombin or Factor Xa protein cleavage signal. Proprotein convertases are a family of nine proteinases, comprising seven basic amino acid-specific subtilisin-like serine proteinases related to yeast kexin, known as prohormone convertase 1/3 (PC1/3), PC2, furin, PC4, PC5/6, paired basic amino-acid cleaving enzyme 4 (PACE4) and PC7, and two other subtilases that cleave at non-basic residues, called subtilisin kexin isozyme 1 (SKI-1) and proprotein convertase subtilisin kexin 9 (PCSK9). Non-limiting examples of protein cleavage signal amino acid sequences are listing in Table 8. In Table 8, “X” refers to any amino acid, “n” may be 0, 2, 4 or 6 amino acids and “*” refers to the protein cleavage site. In Table 8, SEQ ID NO: 2499 refers to when n=4 and SEQ ID NO: 2500 refers to when n=6.

TABLE 8
Protein Cleavage Site Sequences
Protein
CleavageAmino Acid
SignalCleavage SequenceSEQ ID NO
ProproteinR-X-X-R*2497
convertaseR-X-K/R-R*2498
K/R-Xn-K/R*2499 or 2500
ThrombinL-V-P-R*-G-S2501
L-V-P-R*2502
A/F/G/I/L/T/V/M-A/F/G/I/2503
L/T/V/W/A-P-R*
Factor XaI-E-G-R*2504
I-D-G-R*2505
A-E-G-R*2506
A/F/G/I/L/T/V/M-D/E-G-R*2507

In one embodiment, the signal-sensor primary constructs and the mmRNA of the present invention may be engineered such that the primary construct or mmRNA contains at least one encoded protein cleavage signal. The encoded protein cleavage signal may be located before the start codon, after the start codon, before the coding region, within the coding region such as, but not limited to, half way in the coding region, between the start codon and the half way point, between the half way point and the stop codon, after the coding region, before the stop codon, between two stop codons, after the stop codon and combinations thereof.

In one embodiment, the signal-sensor primary constructs or mmRNA of the present invention may include at least one encoded protein cleavage signal containing at least one protein cleavage site. The encoded protein cleavage signal may include, but is not limited to, a proprotein convertase (or prohormone convertase), thrombin and/or Factor Xa protein cleavage signal. One of skill in the art may use Table 1 above or other known methods to determine the appropriate encoded protein cleavage signal to include in the signal-sensor primary constructs or mmRNA of the present invention. For example, starting with the signal of Table 8 and considering the codons of Table 1 one can design a signal for the signal-sensor primary construct which can produce a protein signal in the resulting oncology-related polypeptide.

In one embodiment, the oncology-related polypeptides of the present invention include at least one protein cleavage signal and/or site.

As a non-limiting example, U.S. Pat. No. 7,374,930 and U.S. Pub. No. 20090227660, herein incorporated by reference in their entireties, use a furin cleavage site to cleave the N-terminal methionine of GLP-1 in the expression product from the Golgi apparatus of the cells. In one embodiment, the polypeptides of the present invention include at least one protein cleavage signal and/or site with the proviso that the polypeptide is not GLP-1.

In one embodiment, the signal-sensor primary constructs or mmRNA of the present invention includes at least one encoded protein cleavage signal and/or site.

In one embodiment, the signal-sensor primary constructs or mmRNA of the present invention includes at least one encoded protein cleavage signal and/or site with the proviso that the signal-sensor primary construct or mmRNA does not encode GLP-1.

In one embodiment, the signal-sensor primary constructs or mmRNA of the present invention may include more than one coding region. Where multiple coding regions are present in the signal-sensor primary construct or mmRNA of the present invention, the multiple coding regions may be separated by encoded protein cleavage sites. As a non-limiting example, the signal-sensor primary construct or mmRNA may be signed in an ordered pattern. On such pattern follows AXBY form where A and B are coding regions which may be the same or different coding regions and/or may encode the same or different oncology-related polypeptides, and X and Y are encoded protein cleavage signals which may encode the same or different protein cleavage signals. A second such pattern follows the form AXYBZ where A and B are coding regions which may be the same or different coding regions and/or may encode the same or different oncology-related polypeptides, and X, Y and Z are encoded protein cleavage signals which may encode the same or different protein cleavage signals. A third pattern follows the form ABXCY where A, B and C are coding regions which may be the same or different coding regions and/or may encode the same or different oncology-related polypeptides, and X and Y are encoded protein cleavage signals which may encode the same or different protein cleavage signals.

In one embodiment, the oncology-related polypeptides, signal-sensor primary constructs and mmRNA can also contain sequences that encode protein cleavage sites so that the polypeptides, signal-sensor primary constructs and mmRNA can be released from a carrier region or a fusion partner by treatment with a specific protease for said protein cleavage site.

microRNA

microRNAs (or miRNA) are 19-25 nucleotide long noncoding RNAs that bind to the 3′UTR of nucleic acid molecules and down-regulate gene expression either by reducing nucleic acid molecule stability or by inhibiting translation. The modified nucleic acids (mRNA), enhanced modified RNA or ribonucleic acids of the invention may comprise one or more microRNA target sequences, microRNA sequences, or microRNA seeds. Such sequences may correspond to any known microRNA such as those taught in US Publication US2005/0261218 and US Publication US2005/0059005, the contents of which are incorporated herein by reference in their entirety. As a non-limiting embodiment, known microRNAs, their sequences and their binding site sequences in the human genome are listed below in Table 9.

A microRNA sequence comprises a “seed” region, i.e., a sequence in the region of positions 2-8 of the mature microRNA, which sequence has perfect Watson-Crick complementarity to the miRNA target sequence. A microRNA seed may comprise positions 2-8 or 2-7 of the mature microRNA. In some embodiments, a microRNA seed may comprise 7 nucleotides (e.g., nucleotides 2-8 of the mature microRNA), wherein the seed-complementary site in the corresponding miRNA target is flanked by an adenine (A) opposed to microRNA position 1. In some embodiments, a microRNA seed may comprise 6 nucleotides (e.g., nucleotides 2-7 of the mature microRNA), wherein the seed-complementary site in the corresponding miRNA target is flanked by an adenine (A) opposed to microRNA position 1. See for example, Grimson A, Farh K K, Johnston W K, Garrett-Engele P, Lim L P, Bartel D P; Mol Cell. 2007 Jul. 6; 27(1):91-105. The bases of the microRNA seed have complete complementarity with the target sequence. By engineering microRNA target sequences into the 3′UTR of nucleic acids or mRNA of the invention one can target the molecule for degradation or reduced translation, provided the microRNA in question is available. This process will reduce the hazard of off target effects upon nucleic acid molecule delivery. Identification of microRNA, microRNA target regions, and their expression patterns and role in biology have been reported (Bonauer et al., Curr Drug Targets 2010 11:943-949; Anand and Cheresh Curr Opin Hematol 2011 18:171-176; Contreras and Rao Leukemia 2012 26:404-413 (2011 Dec. 20. doi: 10.1038/leu.2011.356); Bartel Cell 2009 136:215-233; Landgraf et al, Cell, 2007 129:1401-1414; Gentner and Naldini, Tissue Antigens. 2012 80:393-403 and all references therein; each of which is herein incorporated by reference in its entirety).

For example, if the signal-sensor polynucleotide is not intended to be delivered to the liver but ends up there, then miR-122, a microRNA abundant in liver, can inhibit the expression of the gene of interest if one or multiple target sites of miR-122 are engineered into the 3′UTR of the signal-sensor polynucleotide. Introduction of one or multiple binding sites for different microRNA can be engineered to further decrease the longevity, stability, and protein translation of a signal-sensor polynucleotide. As used herein, the term “microRNA site” refers to a microRNA target site or a microRNA recognition site, or any nucleotide sequence to which a microRNA binds or associates. It should be understood that “binding” may follow traditional Watson-Crick hybridization rules or may reflect any stable association of the microRNA with the target sequence at or adjacent to the microRNA site.

Conversely, for the purposes of the signal-sensor polynucleotides of the present invention, microRNA binding sites can be engineered out of (i.e. removed from) sequences in which they naturally occur in order to increase protein expression in specific tissues. For example, miR-122 binding sites may be removed to improve protein expression in the liver.

In one embodiment, signal-sensor polynucleotides may include at least one miRNA-binding site in the 3′UTR in order to direct cytotoxic or cytoprotective mRNA therapeutics to specific cells such as, but not limited to, normal and/or cancerous cells (e.g., HEP3B or SNU449). As a non-limiting example, a strong apoptotic signal and at least one miR-122a binding site is encoded by the signal-sensor polynucleotide where the at least one miR-122a binding site is located in the 3′UTR. As another non-limiting example, apoptosis inducing factor short isoform (AIFsh) and at least one miR-122a binding site is encoded by the signal-sensor polynucleotide where the at least one miR-122a binding site is located in the 3′UTR. As yet another non-limiting example, constitutively active (C.A.) caspase 6 and at least one miR-122a binding site is encoded by the signal-sensor polynucleotide where the at least one miR-122a binding site is located in the 3′UTR. As another non-limiting example, HSV1-tk and at least one miR-122a binding site is encoded by the signal-sensor polynucleotide where the at least one miR-122a binding site is located in the 3′UTR.

In another embodiment, signal-sensor polynucleotides may include three miRNA-binding sites in the 3′UTR in order to direct cytotoxic or cytoprotective mRNA therapeutics to specific cells such as, but not limited to, normal and/or cancerous cells (e.g., HEP3B or SNU449). As a non-limiting example, a strong apoptotic signal and three miR-122a binding sites are encoded by the signal-sensor polynucleotide where the three miR-122a binding sites are located in the 3′UTR. As another non-limiting example, apoptosis inducing factor short isoform (AIFsh) and three miR-122a binding sites are encoded by the signal-sensor polynucleotide where the three miR-122a binding sites are located in the 3′UTR. As yet another non-limiting example, constitutively active (C.A.) caspase 6 and three miR-122a binding sites are encoded by the signal-sensor polynucleotide where the three miR-122a binding sites are located in the 3′UTR. As another non-limiting example, HSV1-tk and three miR-122a binding sites are encoded by the signal-sensor polynucleotide where the three miR-122a binding sites are located in the 3′UTR.

Regulation of expression in multiple tissues can be accomplished through introduction or removal or one or several microRNA binding sites. Shown below in Table 10 are microRNAs which are differentially expressed in different tissues and cells, and often associated with different types of diseases (e.g. cancer cells). The decision of removal or insertion of microRNA binding sites, or any combination, is dependent on microRNA expression patterns and their profilings in cancer cells.

Examples of tissues where microRNA are known to regulate mRNA, and thereby protein expression, include, but are not limited to, liver (miR-122), muscle (miR-133, miR-206, miR-208), endothelial cells (miR-17-92, miR-126), myeloid cells (miR-142-3p, miR-142-5p, miR-16, miR-21, miR-223, miR-24, miR-27), nervous system (mir-124a, miR-9), pluripotent cells (miR-302, miR-367, miR-290, miR-371, miR-373), pancreatic islet cells (miR-375), adipose tissue (let-7, miR-30c), heart (miR-1d, miR-149), kidney (miR-192, miR-194, miR-204), and lung epithelial cells (let-7, miR-133, miR-126).

Specifically, microRNAs are known to be differentially expressed in immune cells (also called hematopoietic cells), such as antigen presenting cells (APCs) (e.g. dendritic cells and macrophages), macrophages, monocytes, B lymphocytes, T lymphocytes, granuocytes, natural killer cells, etc. Immune cell specific microRNAs are involved in immunogenicity, autoimmunity, the immune-response to infection, inflammation, as well as unwanted immune response after gene therapy and tissue/organ transplantation. Immune cells specific microRNAs also regulate many aspects of development, proliferation, differentiation and apoptosis of hematopoietic cells (immune cells). For example, miR-142 and miR-146 are exclusively expressed in the immune cells, particularly abundant in myeloid dendritic cells. Introducing the miR-142 binding site into the 3′-UTR of a signal-sensor polypeptide of the present invention can selectively suppress the gene expression in the antigen presenting cells through miR-142 mediated mRNA degradation, limiting antigen presentation in professional APCs (e.g. dendritic cells) and thereby preventing antigen-mediated immune response after gene delivery (see, Annoni A et al., blood, 2009, 114, 5152-5161, the content of which is herein incorporated by reference in its entirety.)

In one embodiment, microRNAs binding sites that are known to be expressed in immune cells, in particular, the antigen presenting cells, can be engineered into the signal-sensor polynucleotides to suppress the expression of the sensor-signal polynucleotide in APCs through microRNA mediated RNA degradation, subduing the antigen-mediated immune response, while the expression of the sensor-signal polynucleotide is maintained in non-immune cells where the immune cell specific microRNAs are not expressed. For example, to prevent the immunogenic reaction caused by a liver specific protein expression, the miR-122 binding site can be removed and the miR-142 (and/or mirR-146) binding sites can be engineered into the 3-UTR of the signal-sensor polynucleotide (e.g., see the constructs described in Example 38 and the experiment outlined in Examples 39 and 40).

To further drive the selective degradation and suppression of mRNA in APCs and macrophage, the signal-sensor polynucleotide may include another negative regulatory element in the 3-UTR, either alone or in combination with mir-142 and/or mir-146 binding sites. As a non-limiting example, one regulatory element is the Constitutive Decay Elements (CDEs).

Immune cells specific microRNAs include, but are not limited to, hsa-let-7a-2-3p, hsa-let-7a-3p, hsa-7a-5p, hsa-let-7c, hsa-let-7e-3p, hsa-let-7e-5p, hsa-let-7g-3p, hsa-let-7g-5p, hsa-let-7i-3p, hsa-let-7i-5p, miR-10a-3p, miR-10a-5p, miR-1184, hsa-let-7f-1-3p, hsa-let-7f-2-5p, hsa-let-7f-5p, miR-125b-1-3p, miR-125b-2-3p, miR-125b-5p, miR-1279, miR-130a-3p, miR-130a-5p, miR-132-3p, miR-132-5p, miR-142-3p, miR-142-5p, miR-143-3p, miR-143-5p, miR-146a-3p, miR-146a-5p, miR-146b-3p, miR-146b-5p, miR-147a, miR-147b, miR-148a-5p, miR-148a-3p, miR-150-3p, miR-150-5p, miR-151b, miR-155-3p, miR-155-5p, miR-15a-3p, miR-15a-5p, miR-15b-5p, miR-15b-3p, miR-16-1-3p, miR-16-2-3p, miR-16-5p, miR-17-5p, miR-181a-3p, miR-181a-5p, miR-181a-2-3p, miR-182-3p, miR-182-5p, miR-197-3p, miR-197-5p, miR-21-5p, miR-21-3p, miR-214-3p, miR-214-5p, miR-223-3p, miR-223-5p, miR-221-3p, miR-221-5p, miR-23b-3p, miR-23b-5p, miR-24-1-5p, miR-24-2-5p, miR-24-3p, miR-26a-1-3p, miR-26a-2-3p, miR-26a-5p, miR-26b-3p, miR-26b-5p, miR-27a-3p, miR-27a-5p, miR-27b-3p, miR-27b-5p, miR-28-3p, miR-28-5p, miR-2909, miR-29a-3p, miR-29a-5p, miR-29b-1-5p, miR-29b-2-5p, miR-29c-3p, miR-29c-5p, miR-30e-3p, miR-30e-5p, miR-331-5p, miR-339-3p, miR-339-5p, miR-345-3p, miR-345-5p, miR-346, miR-34a-3p, miR-34a-5p, miR-363-3p, miR-363-5p, miR-372, miR-377-3p, miR-377-5p, miR-493-3p, miR-493-5p, miR-542, miR-548b-5p, miR548c-5p, miR-548i, miR-548j, miR-548n, miR-574-3p, miR-598, miR-718, miR-935, miR-99a-3p, miR-99a-5p, miR-99b-3p and miR-99b-5p. Shown below in Table 11 are microRNAs that are enriched in specific types of immune cells. Furthermore, novel microRNAs are discovered in the immune cells in the art through micro-array hybridization and microtome analysis (Jima D D et al, Blood, 2010, 116:e118-e127; Vaz C et al., BMC Genomics, 2010, 11,288, the content of each of which is incorporated herein by reference in its entirety).

MicroRNAs that are known to be expressed in the liver include, but are not limited to, miR-107, miR-122-3p, miR-122-5p, miR-1228-3p, miR-1228-5p, miR-1249, miR-129-5p, miR-1303, miR-151a-3p, miR-151a-5p, miR-152, miR-194-3p, miR-194-5p, miR-199a-3p, miR-199a-5p, miR-199b-3p, miR-199b-5p, miR-296-5p, miR-557, miR-581, miR-939-3p, miR-939-5p. microRNA binding sites from any liver specific microRNA can be introduced to or removed from the signal-sensor polynucleotides to regulate the expression of the signal-sensor polynucleotides in the liver. Liver specific microRNAs binding sites can be engineered alone or further in combination with immune cells (e.g. APCs) microRNA binding sites in order to prevent an immune reaction against protein expression in the liver.

MicroRNAs that are known to be expressed in the lung include, but are not limited to, let-7a-2-3p, let-7a-3p, let-7a-5p, miR-126-3p, miR-126-5p, miR-127-3p, miR-127-5p, miR-130a-3p, miR-130a-5p, miR-130b-3p, miR-130b-5p, miR-133a, miR-133b, miR-134, miR-18a-3p, miR-18a-5p, miR-18b-3p, miR-18b-5p, miR-24-1-5p, miR-24-2-5p, miR-24-3p, miR-296-3p, miR-296-5p, miR-32-3p, miR-337-3p, miR-337-5p, miR-381-3p, miR-381-5p. MicroRNA binding sites from any lung specific microRNA can be introduced to or removed from the signal-sensor polynucleotide to regulate the expression of the signal-sensor polynucleotide in the lung. Lung specific microRNAs binding sites can be engineered alone or further in combination with immune cells (e.g. APCs) microRNA binding sites in order to prevent an immune reaction against protein expression in the lung.

MicroRNAs that are known to be expressed in the heart include, but are not limited to, miR-1, miR-133a, miR-133b, miR-149-3p, miR-149-5p, miR-186-3p, miR-186-5p, miR-208a, miR-208b, miR-210, miR-296-3p, miR-320, miR-451a, miR-451b, miR-499a-3p, miR-499a-5p, miR-499b-3p, miR-499b-5p, miR-744-3p, miR-744-5p, miR-92b-3p and miR-92b-5p. microRNA binding sites from any heart specific microRNA can be introduced to or removed from the signal-sensor polynucleotides to regulate the expression of the signal-sensor polynucleotides in the heart. Heart specific microRNAs binding sites can be engineered alone or further in combination with immune cells (e.g. APCs) microRNA binding sites in order to prevent an immune reaction against protein expression in the heart.

MicroRNAs that are known to be expressed in the nervous system include, but are not limited to, miR-124-5p, miR-125a-3p, miR-125a-5p, miR-125b-1-3p, miR-125b-2-3p, miR-125b-5p, miR-1271-3p, miR-1271-5p, miR-128, miR-132-5p, miR-135a-3p, miR-135a-5p, miR-135b-3p, miR-135b-5p, miR-137, miR-139-5p, miR-139-3p, miR-149-3p, miR-149-5p, miR-153, miR-181c-3p, miR-181c-5p, miR-183-3p, miR-183-5p, miR-190a, miR-190b, miR-212-3p, miR-212-5p, miR-219-1-3p, miR-219-2-3p, miR-23a-3p, miR-23a-5p, miR-30a-5p, miR-30b-3p, miR-30b-5p, miR-30c-1-3p, miR-30c-2-3p, miR-30c-5p, miR-30d-3p, miR-30d-5p, miR-329, miR-342-3p, miR-3665, miR-3666, miR-380-3p, miR-380-5p, miR-383, miR-410, miR-425-3p, miR-425-5p, miR-454-3p, miR-454-5p, miR-483, miR-510, miR-516a-3p, miR-548b-5p, miR-548c-5p, miR-571, miR-7-1-3p, miR-7-2-3p, miR-7-5p, miR-802, miR-922, miR-9-3p and miR-9-5p. microRNAs enriched in the nervous system further include those specifically expressed in neurons, including, but not limited to, miR-132-3p, miR-132-3p, miR-148b-3p, miR-148b-5p, miR-151a-3p, miR-151a-5p, miR-212-3p, miR-212-5p, miR-320b, miR-320e, miR-323a-3p, miR-323a-5p, miR-324-5p, miR-325, miR-326, miR-328, miR-922 and those specifically expressed in glial cells, including, but not limited to, miR-1250, miR-219-1-3p, miR-219-2-3p, miR-219-5p, miR-23a-3p, miR-23a-5p, miR-3065-3p, miR-3065-5p, miR-30e-3p, miR-30e-5p, miR-32-5p, miR-338-5p, miR-657. microRNA binding sites from any CNS specific microRNA can be introduced to or removed from the signal-sensor polynucleotides to regulate the expression of the signal-sensor polynucleotide in the nervous system. Nervous system specific microRNAs binding sites can be engineered alone or further in combination with immune cells (e.g. APCs) microRNA binding sites in order to prevent an immune reaction against protein expression in the nervous system.

MicroRNAs that are known to be expressed in the pancreas include, but are not limited to, miR-105-3p, miR-105-5p, miR-184, miR-195-3p, miR-195-5p, miR-196a-3p, miR-196a-5p, miR-214-3p, miR-214-5p, miR-216a-3p, miR-216a-5p, miR-30a-3p, miR-33a-3p, miR-33a-5p, miR-375, miR-7-1-3p, miR-7-2-3p, miR-493-3p, miR-493-5p and miR-944. MicroRNA binding sites from any pancreas specific microRNA can be introduced to or removed from the signal-sensor polynucleotide to regulate the expression of the signal-sensor polynucleotide in the pancreas. Pancreas specific microRNAs binding sites can be engineered alone or further in combination with immune cells (e.g. APCs) microRNA binding sites in order to prevent immune reaction against protein expression in the pancreas.

MicroRNAs that are known to be expressed in the kidney further include, but are not limited to, miR-122-3p, miR-145-5p, miR-17-5p, miR-192-3p, miR-192-5p, miR-194-3p, miR-194-5p, miR-20a-3p, miR-20a-5p, miR-204-3p, miR-204-5p, miR-210, miR-216a-3p, miR-216a-5p, miR-296-3p, miR-30a-3p, miR-30a-5p, miR-30b-3p, miR-30b-5p, miR-30c-1-3p, miR-30c-2-3p, miR30c-5p, miR-324-3p, miR-335-3p, miR-335-5p, miR-363-3p, miR-363-5p and miR-562. MicroRNA binding sites from any kidney specific microRNA can be introduced to or removed from the signal-sensor polynucleotide to regulate the expression of the signal-sensor polynucleotide in the kidney. Kidney specific microRNAs binding sites can be engineered alone or further in combination with immune cells (e.g. APCs) microRNA binding sites in order to prevent immune reaction against protein expression in the kidney.

MicroRNAs that are known to be expressed in the muscle further include, but are not limited to, let-7g-3p, let-7g-5p, miR-1, miR-1286, miR-133a, miR-133b, miR-140-3p, miR-143-3p, miR-143-5p, miR-145-3p, miR-145-5p, miR-188-3p, miR-188-5p, miR-206, miR-208a, miR-208b, miR-25-3p and miR-25-5p. MicroRNA binding sites from any muscle specific microRNA can be introduced to or removed from the signal-sensor polynucleotide to regulate the expression of the signal-sensor polynucleotide in the muscle. Muscle specific microRNAs binding sites can be engineered alone or further in combination with immune cells (e.g. APCs) microRNA binding sites in order to prevent an immune reaction against protein expression in the muscle.

MicroRNAs are differentially expressed in different types of cells, such as endothelial cells, epithelial cells and adipocytes. For example, microRNAs that are expressed in endothelial cells include, but are not limited to, let-7b-3p, let-7b-5p, miR-100-3p, miR-100-5p, miR-101-3p, miR-101-5p, miR-126-3p, miR-126-5p, miR-1236-3p, miR-1236-5p, miR-130a-3p, miR-130a-5p, miR-17-5p, miR-17-3p, miR-18a-3p, miR-18a-5p, miR-19a-3p, miR-19a-5p, miR-19b-1-5p, miR-19b-2-5p, miR-19b-3p, miR-20a-3p, miR-20a-5p, miR-217, miR-210, miR-21-3p, miR-21-5p, miR-221-3p, miR-221-5p, miR-222-3p, miR-222-5p, miR-23a-3p, miR-23a-5p, miR-296-5p, miR-361-3p, miR-361-5p, miR-421, miR-424-3p, miR-424-5p, miR-513a-5p, miR-92a-1-5p, miR-92a-2-5p, miR-92a-3p, miR-92b-3p and miR-92b-5p. Many novel microRNAs were discovered in endothelial cells from deep-sequencing analysis (Voellenkle C et al., RNA, 2012, 18, 472-484, herein incorporated by reference in its entirety). MicroRNA binding sites from any endothelial cell specific microRNA can be introduced to or removed from the signal-sensor polynucleotide in order to modulate the expression of the signal-sensor polynucleotide in the endothelial cells in various conditions.

For further example, microRNAs that are expressed in epithelial cells include, but are not limited to, let-7b-3p, let-7b-5p, miR-1246, miR-200a-3p, miR-200a-5p, miR-200b-3p, miR-200b-5p, miR-200c-3p, miR-200c-5p, miR-338-3p, miR-429, miR-451a, miR-451b, miR-494, miR-802 and miR-34a, miR-34b-5p, miR-34c-5p, miR-449a, miR-449b-3p, miR-449b-5p specific in respiratory ciliated epithelial cells; let-7 family, miR-133a, miR-133b, miR-126 specific in lung epithelial cells; miR-382-3p, miR-382-5p specific in renal epithelial cells and miR-762 specific in corneal epithelial cells. MicroRNA binding sites from any epithelial cell specific microRNA can be introduced to or removed from the signal-sensor polynucleotide in order to modulate the expression of the signal-sensor polynucleotide in the epithelial cells in various conditions.

In addition, a large group of microRNAs are enriched in embryonic stem cells, controlling stem cell self-renewal as well as the development and/or differentiation of various cell lineages, such as neural cells, cardiac, hematopoietic cells, skin cells, osteogenic cells and muscle cells (Kuppusamy K T et al., Curr. Mol Med, 2013, 13(5), 757-764; Vidigal J A and Ventura A, Semin Cancer Biol. 2012, 22(5-6), 428-436; Goff L A et al., PLoS One, 2009, 4:e7192; Morin R D et al., Genome Res, 2008, 18, 610-621; Yoo J K et al., Stem Cells Dev. 2012, 21(11), 2049-2057, each of which is herein incorporated by reference in its entirety). MicroRNAs abundant in embryonic stem cells include, but are not limited to, let-7a-2-3p, let-a-3p, let-7a-5p, let7d-3p, let-7d-5p, miR-103a-2-3p, miR-103a-5p, miR-106b-3p, miR-106b-5p, miR-1246, miR-1275, miR-138-1-3p, miR-138-2-3p, miR-138-5p, miR-154-3p, miR-154-5p, miR-200c-3p, miR-200c-5p, miR-290, miR-301a-3p, miR-301a-5p, miR-302a-3p, miR-302a-5p, miR-302b-3p, miR-302b-5p, miR-302c-3p, miR-302c-5p, miR-302d-3p, miR-302d-5p, miR-302e, miR-367-3p, miR-367-5p, miR-369-3p, miR-369-5p, miR-370, miR-371, miR-373, miR-380-5p, miR-423-3p, miR-423-5p, miR-486-5p, miR-520c-3p, miR-548e, miR-548f, miR-548g-3p, miR-548g-5p, miR-548i, miR-548k, miR-548l, miR-548m, miR-548n, miR-548o-3p, miR-548o-5p, miR-548p, miR-664a-3p, miR-664a-5p, miR-664b-3p, miR-664b-5p, miR-766-3p, miR-766-5p, miR-885-3p, miR-885-5p, miR-93-3p, miR-93-5p, miR-941, miR-96-3p, miR-96-5p, miR-99b-3p and miR-99b-5p. Many predicted novel microRNAs are discovered by deep sequencing in human embryonic stem cells (Morin R D et al., Genome Res, 2008, 18, 610-621; Goff L A et al., PLoS One, 2009, 4:e7192; Bar M et al., Stem cells, 2008, 26, 2496-2505, the content of each of which is incorporated herein by references in its entirety).

In one embodiment, the binding sites of embryonic stem cell specific microRNAs can be included in or removed from the 3-UTR of the signal-sensor polynucleotide to modulate the development and/or differentiation of embryonic stem cells, to inhibit the senescence of stem cells in a degenerative condition (e.g. degenerative diseases), or to stimulate the senescence and apoptosis of stem cells in a disease condition (e.g. cancer stem cell).

Many microRNA expression studies have been conducted, and are described in the art, to profile the differential expression of microRNAs in various cancer cells/tissues and other diseases. Some microRNAs are abnormally over-expressed in certain cancer cells and others are under-expressed. For example, microRNAs are differentially expressed in cancer cells (WO2008/154098, US2013/0059015, US2013/0042333, WO2011/157294); cancer stem cells (US2012/0053224); pancreatic cancers and diseases (US2009/0131348, US2011/0171646, US2010/0286232, U.S. Pat. No. 8,389,210); asthma and inflammation (U.S. Pat. No. 8,415,096); prostate cancer (US2013/0053264); hepatocellular carcinoma (WO2012/151212, US2012/0329672, WO2008/054828, U.S. Pat. No. 8,252,538); lung cancer cells (WO2011/076143, WO2013/033640, WO2009/070653, US2010/0323357); cutaneous T cell lymphoma (WO2013/011378); colorectal cancer cells (WO2011/0281756, WO2011/076142); cancer positive lympho nodes (WO2009/100430, US2009/0263803); nasopharyngeal carcinoma (EP2112235); chronic obstructive pulmonary disease (US2012/0264626, US2013/0053263); thyroid cancer (WO2013/066678); ovarian cancer cells (US2012/0309645, WO2011/095623); breast cancer cells (WO2008/154098, WO2007/081740, US2012/0214699), leukemia and lymphoma (WO2008/073915, US2009/0092974, US2012/0316081, US2012/0283310, WO2010/018563, the content of each of which is incorporated herein by reference in their entirety).

Specifically, microRNA sites that are over-expressed in certain cancer and/or tumor cells can be removed from the 3-UTR of the signal-sensor polynucleotide encoding the oncology-related polypeptide, restoring the expression suppressed by the over-expressed microRNAs in cancer cells, thus ameliorating the corresponsive biological function, for instance, transcription stimulation and/or repression, cell cycle arrest, apoptosis and cell death. Normal cells and tissues, wherein microRNA expression is not up-regulated, will remain unaffected.

MicroRNA can also regulate complex biological processes such as angiogenesis (miR-132) (Anand and Cheresh Curr Opin Hematol 2011 18:171-176). In the signal-sensor polynucleotides of the invention, binding sites for microRNAs that are involved in such processes may be removed or introduced, in order to tailor the expression of the signal-sensor polynucleotides expression to biologically relevant cell types or to the context of relevant biological processes. In this context, the signal-sensor polynucleotides are defined as auxotrophic signal-sensor polynucleotides.

Table 9 is a non-exhaustive listing of miRs and miR binding sites (miR BS) and their sequences which may be used with the present invention.

TABLE 9
Mirs and mir binding sites
mir SEQBS SEQ
microRNAIDID
hsa-let-7a-2-3p25083529
hsa-let-7a-3p25093530
hsa-let-7a-5p25103531
hsa-let-7b-3p25113532
hsa-let-7b-5p25123533
hsa-let-7c25133534
hsa-let-7d-3p25143535
hsa-let-7d-5p25153536
hsa-let-7e-3p25163537
hsa-let-7e-5p25173538
hsa-let-7f-1-3p25183539
hsa-let-7f-2-3p25193540
hsa-let-7f-5p25203541
hsa-let-7g-3p25213542
hsa-let-7g-5p25223543
hsa-let-7i-3p25233544
hsa-let-7i-5p25243545
hsa-miR-125253546
hsa-miR-100-3p25263547
hsa-miR-100-5p25273548
hsa-miR-101-3p25283549
hsa-miR-101-5p25293550
hsa-miR-103a-2-5p25303551
hsa-miR-103a-3p25313552
hsa-miR-103b25323553
hsa-miR-105-3p25333554
hsa-miR-105-5p25343555
hsa-miR-106a-3p25353556
hsa-miR-106a-5p25363557
hsa-miR-106b-3p25373558
hsa-miR-106b-5p25383559
hsa-miR-10725393560
hsa-miR-10a-3p25403561
hsa-miR-10a-5p25413562
hsa-miR-10b-3p25423563
hsa-miR-10b-5p25433564
hsa-miR-1178-3p25443565
hsa-miR-1178-5p25453566
hsa-miR-117925463567
hsa-miR-118025473568
hsa-miR-118125483569
hsa-miR-118225493570
hsa-miR-118325503571
hsa-miR-118425513572
hsa-miR-1185-1-3p25523573
hsa-miR-1185-2-3p25533574
hsa-miR-1185-5p25543575
hsa-miR-119325553576
hsa-miR-119725563577
hsa-miR-120025573578
hsa-miR-120225583579
hsa-miR-120325593580
hsa-miR-120425603581
hsa-miR-120525613582
hsa-miR-120625623583
hsa-miR-1207-3p25633584
hsa-miR-1207-5p25643585
hsa-miR-120825653586
hsa-miR-122-3p25663587
hsa-miR-1224-3p25673588
hsa-miR-1224-5p25683589
hsa-miR-1225-3p25693590
hsa-miR-1225-5p25703591
hsa-miR-122-5p25713592
hsa-miR-1226-3p25723593
hsa-miR-1226-5p25733594
hsa-miR-1227-3p25743595
hsa-miR-1227-5p25753596
hsa-miR-1228-3p25763597
hsa-miR-1228-5p25773598
hsa-miR-1229-3p25783599
hsa-miR-1229-5p25793600
hsa-miR-123125803601
hsa-miR-1233-1-5p25813602
hsa-miR-1233-3p25823603
hsa-miR-1234-3p25833604
hsa-miR-1234-5p25843605
hsa-miR-1236-3p25853606
hsa-miR-1236-5p25863607
hsa-miR-1237-3p25873608
hsa-miR-1237-5p25883609
hsa-miR-1238-3p25893610
hsa-miR-1238-5p25903611
hsa-miR-124325913612
hsa-miR-124-3p25923613
hsa-miR-124425933614
hsa-miR-1245a25943615
hsa-miR-1245b-3p25953616
hsa-miR-1245b-5p25963617
hsa-miR-124-5p25973618
hsa-miR-124625983619
hsa-miR-1247-3p25993620
hsa-miR-1247-5p26003621
hsa-miR-124826013622
hsa-miR-124926023623
hsa-miR-125026033624
hsa-miR-125126043625
hsa-miR-125226053626
hsa-miR-125326063627
hsa-miR-125426073628
hsa-miR-1255a26083629
hsa-miR-1255b-2-3p26093630
hsa-miR-1255b-5p26103631
hsa-miR-125626113632
hsa-miR-125726123633
hsa-miR-125826133634
hsa-miR-125a-3p26143635
hsa-miR-125a-5p26153636
hsa-miR-125b-1-3p26163637
hsa-miR-125b-2-3p26173638
hsa-miR-125b-5p26183639
hsa-miR-1260a26193640
hsa-miR-1260b26203641
hsa-miR-126126213642
hsa-miR-126226223643
hsa-miR-126326233644
hsa-miR-126-3p26243645
hsa-miR-126426253646
hsa-miR-126526263647
hsa-miR-126-5p26273648
hsa-miR-126626283649
hsa-miR-126726293650
hsa-miR-1268a26303651
hsa-miR-1268b26313652
hsa-miR-1269a26323653
hsa-miR-1269b26333654
hsa-miR-127026343655
hsa-miR-1271-3p26353656
hsa-miR-1271-5p26363657
hsa-miR-127226373658
hsa-miR-1273a26383659
hsa-miR-1273c26393660
hsa-miR-1273d26403661
hsa-miR-1273e26413662
hsa-miR-1273f26423663
hsa-miR-1273g-3p26433664
hsa-miR-1273g-5p26443665
hsa-miR-127-3p26453666
hsa-miR-127526463667
hsa-miR-127-5p26473668
hsa-miR-127626483669
hsa-miR-1277-3p26493670
hsa-miR-1277-5p26503671
hsa-miR-127826513672
hsa-miR-127926523673
hsa-miR-12826533674
hsa-miR-128126543675
hsa-miR-128226553676
hsa-miR-128326563677
hsa-miR-128426573678
hsa-miR-1285-3p26583679
hsa-miR-1285-5p26593680
hsa-miR-128626603681
hsa-miR-128726613682
hsa-miR-128826623683
hsa-miR-128926633684
hsa-miR-129026643685
hsa-miR-129126653686
hsa-miR-129-1-3p26663687
hsa-miR-1292-3p26673688
hsa-miR-129-2-3p26683689
hsa-miR-1292-5p26693690
hsa-miR-129326703691
hsa-miR-129426713692
hsa-miR-1295a26723693
hsa-miR-1295b-3p26733694
hsa-miR-1295b-5p26743695
hsa-miR-129-5p26753696
hsa-miR-129626763697
hsa-miR-129726773698
hsa-miR-129826783699
hsa-miR-129926793700
hsa-miR-130126803701
hsa-miR-130226813702
hsa-miR-130326823703
hsa-miR-1304-3p26833704
hsa-miR-1304-5p26843705
hsa-miR-130526853706
hsa-miR-1306-3p26863707
hsa-miR-1306-5p26873708
hsa-miR-1307-3p26883709
hsa-miR-1307-5p26893710
hsa-miR-130a-3p26903711
hsa-miR-130a-5p26913712
hsa-miR-130b-3p26923713
hsa-miR-130b-5p26933714
hsa-miR-132126943715
hsa-miR-132226953716
hsa-miR-132326963717
hsa-miR-132-3p26973718
hsa-miR-132426983719
hsa-miR-132-5p26993720
hsa-miR-133a27003721
hsa-miR-133b27013722
hsa-miR-13427023723
hsa-miR-134327033724
hsa-miR-135a-3p27043725
hsa-miR-135a-5p27053726
hsa-miR-135b-3p27063727
hsa-miR-135b-5p27073728
hsa-miR-136-3p27083729
hsa-miR-136-5p27093730
hsa-miR-13727103731
hsa-miR-138-1-3p27113732
hsa-miR-138-2-3p27123733
hsa-miR-138-5p27133734
hsa-miR-139-3p27143735
hsa-miR-139-5p27153736
hsa-miR-140-3p27163737
hsa-miR-140-5p27173738
hsa-miR-141-3p27183739
hsa-miR-141-5p27193740
hsa-miR-142-3p27203741
hsa-miR-142-5p27213742
hsa-miR-143-3p27223743
hsa-miR-143-5p27233744
hsa-miR-144-3p27243745
hsa-miR-144-5p27253746
hsa-miR-145-3p27263747
hsa-miR-145-5p27273748
hsa-miR-146827283749
hsa-miR-146927293750
hsa-miR-146a-3p27303751
hsa-miR-146a-5p27313752
hsa-miR-146b-3p27323753
hsa-miR-146b-5p27333754
hsa-miR-147027343755
hsa-miR-147127353756
hsa-miR-147a27363757
hsa-miR-147b27373758
hsa-miR-148a-3p27383759
hsa-miR-148a-5p27393760
hsa-miR-148b-3p27403761
hsa-miR-148b-5p27413762
hsa-miR-149-3p27423763
hsa-miR-149-5p27433764
hsa-miR-150-3p27443765
hsa-miR-150-5p27453766
hsa-miR-151a-3p27463767
hsa-miR-151a-5p27473768
hsa-miR-151b27483769
hsa-miR-15227493770
hsa-miR-15327503771
hsa-miR-153727513772
hsa-miR-153827523773
hsa-miR-153927533774
hsa-miR-154-3p27543775
hsa-miR-154-5p27553776
hsa-miR-155-3p27563777
hsa-miR-155-5p27573778
hsa-miR-158727583779
hsa-miR-15a-3p27593780
hsa-miR-15a-5p27603781
hsa-miR-15b-3p27613782
hsa-miR-15b-5p27623783
hsa-miR-16-1-3p27633784
hsa-miR-16-2-3p27643785
hsa-miR-16-5p27653786
hsa-miR-17-3p27663787
hsa-miR-17-5p27673788
hsa-miR-181a-2-3p27683789
hsa-miR-181a-3p27693790
hsa-miR-181a-5p27703791
hsa-miR-181b-3p27713792
hsa-miR-181b-5p27723793
hsa-miR-181c-3p27733794
hsa-miR-181c-5p27743795
hsa-miR-181d27753796
hsa-miR-182-3p27763797
hsa-miR-182527773798
hsa-miR-182-5p27783799
hsa-miR-182727793800
hsa-miR-183-3p27803801
hsa-miR-183-5p27813802
hsa-miR-18427823803
hsa-miR-185-3p27833804
hsa-miR-185-5p27843805
hsa-miR-186-3p27853806
hsa-miR-186-5p27863807
hsa-miR-187-3p27873808
hsa-miR-187-5p27883809
hsa-miR-188-3p27893810
hsa-miR-188-5p27903811
hsa-miR-18a-3p27913812
hsa-miR-18a-5p27923813
hsa-miR-18b-3p27933814
hsa-miR-18b-5p27943815
hsa-miR-190827953816
hsa-miR-1909-3p27963817
hsa-miR-1909-5p27973818
hsa-miR-190a27983819
hsa-miR-190b27993820
hsa-miR-191028003821
hsa-miR-1911-3p28013822
hsa-miR-1911-5p28023823
hsa-miR-191228033824
hsa-miR-191328043825
hsa-miR-191-3p28053826
hsa-miR-1914-3p28063827
hsa-miR-1914-5p28073828
hsa-miR-1915-3p28083829
hsa-miR-1915-5p28093830
hsa-miR-191-5p28103831
hsa-miR-192-3p28113832
hsa-miR-192-5p28123833
hsa-miR-193a-3p28133834
hsa-miR-193a-5p28143835
hsa-miR-193b-3p28153836
hsa-miR-193b-5p28163837
hsa-miR-194-3p28173838
hsa-miR-194-5p28183839
hsa-miR-195-3p28193840
hsa-miR-195-5p28203841
hsa-miR-196a-3p28213842
hsa-miR-196a-5p28223843
hsa-miR-196b-3p28233844
hsa-miR-196b-5p28243845
hsa-miR-197228253846
hsa-miR-197328263847
hsa-miR-197-3p28273848
hsa-miR-197-5p28283849
hsa-miR-197628293850
hsa-miR-19828303851
hsa-miR-199a-3p28313852
hsa-miR-199a-5p28323853
hsa-miR-199b-3p28333854
hsa-miR-199b-5p28343855
hsa-miR-19a-3p28353856
hsa-miR-19a-5p28363857
hsa-miR-19b-1-5p28373858
hsa-miR-19b-2-5p28383859
hsa-miR-19b-3p28393860
hsa-miR-200a-3p28403861
hsa-miR-200a-5p28413862
hsa-miR-200b-3p28423863
hsa-miR-200b-5p28433864
hsa-miR-200c-3p28443865
hsa-miR-200c-5p28453866
hsa-miR-202-3p28463867
hsa-miR-202-5p28473868
hsa-miR-203a28483869
hsa-miR-203b-3p28493870
hsa-miR-203b-5p28503871
hsa-miR-204-3p28513872
hsa-miR-204-5p28523873
hsa-miR-205228533874
hsa-miR-205328543875
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hsa-miR-205428563877
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hsa-miR-20628583879
hsa-miR-208a28593880
hsa-miR-208b28603881
hsa-miR-20a-3p28613882
hsa-miR-20a-5p28623883
hsa-miR-20b-3p28633884
hsa-miR-20b-5p28643885
hsa-miR-21028653886
hsa-miR-211028663887
hsa-miR-211328673888
hsa-miR-211-3p28683889
hsa-miR-2114-3p28693890
hsa-miR-2114-5p28703891
hsa-miR-2115-3p28713892
hsa-miR-2115-5p28723893
hsa-miR-211-5p28733894
hsa-miR-2116-3p28743895
hsa-miR-2116-5p28753896
hsa-miR-211728763897
hsa-miR-212-3p28773898
hsa-miR-212-5p28783899
hsa-miR-21-3p28793900
hsa-miR-214-3p28803901
hsa-miR-214-5p28813902
hsa-miR-21528823903
hsa-miR-21-5p28833904
hsa-miR-216a-3p28843905
hsa-miR-216a-5p28853906
hsa-miR-216b28863907
hsa-miR-21728873908
hsa-miR-218-1-3p28883909
hsa-miR-218-2-3p28893910
hsa-miR-218-5p28903911
hsa-miR-219-1-3p28913912
hsa-miR-219-2-3p28923913
hsa-miR-219-5p28933914
hsa-miR-221-3p28943915
hsa-miR-221-5p28953916
hsa-miR-222-3p28963917
hsa-miR-222-5p28973918
hsa-miR-223-3p28983919
hsa-miR-223-5p28993920
hsa-miR-22-3p29003921
hsa-miR-224-3p29013922
hsa-miR-224-5p29023923
hsa-miR-22-5p29033924
hsa-miR-227629043925
hsa-miR-2277-3p29053926
hsa-miR-2277-5p29063927
hsa-miR-227829073928
hsa-miR-2355-3p29083929
hsa-miR-2355-5p29093930
hsa-miR-239229103931
hsa-miR-23a-3p29113932
hsa-miR-23a-5p29123933
hsa-miR-23b-3p29133934
hsa-miR-23b-5p29143935
hsa-miR-23c29153936
hsa-miR-24-1-5p29163937
hsa-miR-24-2-5p29173938
hsa-miR-24-3p29183939
hsa-miR-2467-3p29193940
hsa-miR-2467-5p29203941
hsa-miR-25-3p29213942
hsa-miR-25-5p29223943
hsa-miR-2681-3p29233944
hsa-miR-2681-5p29243945
hsa-miR-2682-3p29253946
hsa-miR-2682-5p29263947
hsa-miR-26a-1-3p29273948
hsa-miR-26a-2-3p29283949
hsa-miR-26a-5p29293950
hsa-miR-26b-3p29303951
hsa-miR-26b-5p29313952
hsa-miR-27a-3p29323953
hsa-miR-27a-5p29333954
hsa-miR-27b-3p29343955
hsa-miR-27b-5p29353956
hsa-miR-28-3p29363957
hsa-miR-28-5p29373958
hsa-miR-286129383959
hsa-miR-290929393960
hsa-miR-296-3p29403961
hsa-miR-2964a-3p29413962
hsa-miR-2964a-5p29423963
hsa-miR-296-5p29433964
hsa-miR-29729443965
hsa-miR-29829453966
hsa-miR-299-3p29463967
hsa-miR-299-5p29473968
hsa-miR-29a-3p29483969
hsa-miR-29a-5p29493970
hsa-miR-29b-1-5p29503971
hsa-miR-29b-2-5p29513972
hsa-miR-29b-3p29523973
hsa-miR-29c-3p29533974
hsa-miR-29c-5p29543975
hsa-miR-30029553976
hsa-miR-301a-3p29563977
hsa-miR-301a-5p29573978
hsa-miR-301b29583979
hsa-miR-302a-3p29593980
hsa-miR-302a-5p29603981
hsa-miR-302b-3p29613982
hsa-miR-302b-5p29623983
hsa-miR-302c-3p29633984
hsa-miR-302c-5p29643985
hsa-miR-302d-3p29653986
hsa-miR-302d-5p29663987
hsa-miR-302e29673988
hsa-miR-302f29683989
hsa-miR-3064-3p29693990
hsa-miR-3064-5p29703991
hsa-miR-3065-3p29713992
hsa-miR-3065-5p29723993
hsa-miR-3074-3p29733994
hsa-miR-3074-5p29743995
hsa-miR-30a-3p29753996
hsa-miR-30a-5p29763997
hsa-miR-30b-3p29773998
hsa-miR-30b-5p29783999
hsa-miR-30c-1-3p29794000
hsa-miR-30c-2-3p29804001
hsa-miR-30c-5p29814002
hsa-miR-30d-3p29824003
hsa-miR-30d-5p29834004
hsa-miR-30e-3p29844005
hsa-miR-30e-5p29854006
hsa-miR-311529864007
hsa-miR-311629874008
hsa-miR-3117-3p29884009
hsa-miR-3117-5p29894010
hsa-miR-311829904011
hsa-miR-311929914012
hsa-miR-3120-3p29924013
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hsa-miR-3121-3p29944015
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hsa-miR-312229964017
hsa-miR-312329974018
hsa-miR-3124-3p29984019
hsa-miR-3124-5p29994020
hsa-miR-312530004021
hsa-miR-3126-3p30014022
hsa-miR-3126-5p30024023
hsa-miR-3127-3p30034024
hsa-miR-3127-5p30044025
hsa-miR-312830054026
hsa-miR-3129-3p30064027
hsa-miR-3129-5p30074028
hsa-miR-3130-3p30084029
hsa-miR-3130-5p30094030
hsa-miR-313130104031
hsa-miR-313230114032
hsa-miR-313330124033
hsa-miR-313430134034
hsa-miR-3135a30144035
hsa-miR-3135b30154036
hsa-miR-3136-3p30164037
hsa-miR-3136-5p30174038
hsa-miR-313730184039
hsa-miR-313830194040
hsa-miR-313930204041
hsa-miR-31-3p30214042
hsa-miR-3140-3p30224043
hsa-miR-3140-5p30234044
hsa-miR-314130244045
hsa-miR-314230254046
hsa-miR-314330264047
hsa-miR-3144-3p30274048
hsa-miR-3144-5p30284049
hsa-miR-3145-3p30294050
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hsa-miR-314630314052
hsa-miR-314730324053
hsa-miR-314830334054
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hsa-miR-3150a-3p30354056
hsa-miR-3150a-5p30364057
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hsa-miR-3150b-5p30384059
hsa-miR-315130394060
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hsa-miR-3152-5p30414062
hsa-miR-315330424063
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hsa-miR-3156-5p30474068
hsa-miR-3157-3p30484069
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hsa-miR-3158-3p30504071
hsa-miR-3158-5p30514072
hsa-miR-315930524073
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hsa-miR-316130564077
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hsa-miR-319230974118
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hsa-miR-319531014122
hsa-miR-319631024123
hsa-miR-319731034124
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hsa-miR-320131084129
hsa-miR-320231094130
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hsa-miR-32-3p31194140
hsa-miR-324-3p31204141
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hsa-miR-34631494170
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hsa-miR-3605-3p31604181
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hsa-miR-360931664187
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hsa-miR-3616-3p31774198
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hsa-miR-3617-3p31794200
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hsa-miR-361831814202
hsa-miR-3619-3p31824203
hsa-miR-3619-5p31834204
hsa-miR-3620-3p31844205
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hsa-miR-4733-3p48135834
hsa-miR-4733-5p48145835
hsa-miR-473448155836
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hsa-miR-473648185839
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hsa-miR-64754036424
hsa-miR-64854046425
hsa-miR-64954056426
hsa-miR-6499-3p54066427
hsa-miR-6499-5p54076428
hsa-miR-65054086429
hsa-miR-6500-3p54096430
hsa-miR-6500-5p54106431
hsa-miR-6501-3p54116432
hsa-miR-6501-5p54126433
hsa-miR-6502-3p54136434
hsa-miR-6502-5p54146435
hsa-miR-6503-3p54156436
hsa-miR-6503-5p54166437
hsa-miR-6504-3p54176438
hsa-miR-6504-5p54186439
hsa-miR-6505-3p54196440
hsa-miR-6505-5p54206441
hsa-miR-6506-3p54216442
hsa-miR-6506-5p54226443
hsa-miR-6507-3p54236444
hsa-miR-6507-5p54246445
hsa-miR-6508-3p54256446
hsa-miR-6508-5p54266447
hsa-miR-6509-3p54276448
hsa-miR-6509-5p54286449
hsa-miR-65154296450
hsa-miR-6510-3p54306451
hsa-miR-6510-5p54316452
hsa-miR-6511a-3p54326453
hsa-miR-6511a-5p54336454
hsa-miR-6511b-3p54346455
hsa-miR-6511b-5p54356456
hsa-miR-6512-3p54366457
hsa-miR-6512-5p54376458
hsa-miR-6513-3p54386459
hsa-miR-6513-5p54396460
hsa-miR-6514-3p54406461
hsa-miR-6514-5p54416462
hsa-miR-6515-3p54426463
hsa-miR-6515-5p54436464
hsa-miR-652-3p54446465
hsa-miR-652-5p54456466
hsa-miR-65354466467
hsa-miR-654-3p54476468
hsa-miR-654-5p54486469
hsa-miR-65554496470
hsa-miR-65654506471
hsa-miR-65754516472
hsa-miR-65854526473
hsa-miR-659-3p54536474
hsa-miR-659-5p54546475
hsa-miR-660-3p54556476
hsa-miR-660-5p54566477
hsa-miR-66154576478
hsa-miR-66254586479
hsa-miR-663a54596480
hsa-miR-663b54606481
hsa-miR-664a-3p54616482
hsa-miR-664a-5p54626483
hsa-miR-664b-3p54636484
hsa-miR-664b-5p54646485
hsa-miR-66554656486
hsa-miR-66854666487
hsa-miR-67054676488
hsa-miR-671-3p54686489
hsa-miR-6715a-3p54696490
hsa-miR-6715b-3p54706491
hsa-miR-6715b-5p54716492
hsa-miR-671-5p54726493
hsa-miR-6716-3p54736494
hsa-miR-6716-5p54746495
hsa-miR-6717-5p54756496
hsa-miR-6718-5p54766497
hsa-miR-6719-3p54776498
hsa-miR-6720-3p54786499
hsa-miR-6721-5p54796500
hsa-miR-6722-3p54806501
hsa-miR-6722-5p54816502
hsa-miR-6723-5p54826503
hsa-miR-6724-5p54836504
hsa-miR-675-3p54846505
hsa-miR-675-5p54856506
hsa-miR-676-3p54866507
hsa-miR-676-5p54876508
hsa-miR-708-3p54886509
hsa-miR-708-5p54896510
hsa-miR-71154906511
hsa-miR-7-1-3p54916512
hsa-miR-71854926513
hsa-miR-7-2-3p54936514
hsa-miR-744-3p54946515
hsa-miR-744-5p54956516
hsa-miR-758-3p54966517
hsa-miR-758-5p54976518
hsa-miR-75954986519
hsa-miR-7-5p54996520
hsa-miR-76055006521
hsa-miR-76155016522
hsa-miR-76255026523
hsa-miR-76455036524
hsa-miR-76555046525
hsa-miR-766-3p55056526
hsa-miR-766-5p55066527
hsa-miR-767-3p55076528
hsa-miR-767-5p55086529
hsa-miR-769-3p55096530
hsa-miR-769-5p55106531
hsa-miR-770-5p55116532
hsa-miR-80255126533
hsa-miR-873-3p55136534
hsa-miR-873-5p55146535
hsa-miR-87455156536
hsa-miR-875-3p55166537
hsa-miR-875-5p55176538
hsa-miR-876-3p55186539
hsa-miR-876-5p55196540
hsa-miR-877-3p55206541
hsa-miR-877-5p55216542
hsa-miR-885-3p55226543
hsa-miR-885-5p55236544
hsa-miR-88755246545
hsa-miR-888-3p55256546
hsa-miR-888-5p55266547
hsa-miR-88955276548
hsa-miR-89055286549
hsa-miR-891a55296550
hsa-miR-891b55306551
hsa-miR-892a55316552
hsa-miR-892b55326553
hsa-miR-892c-3p55336554
hsa-miR-892c-5p55346555
hsa-miR-92055356556
hsa-miR-92155366557
hsa-miR-92255376558
hsa-miR-92455386559
hsa-miR-92a-1-5p55396560
hsa-miR-92a-2-5p55406561
hsa-miR-92a-3p55416562
hsa-miR-92b-3p55426563
hsa-miR-92b-5p55436564
hsa-miR-93355446565
hsa-miR-93-3p55456566
hsa-miR-93455466567
hsa-miR-93555476568
hsa-miR-93-5p55486569
hsa-miR-93655496570
hsa-miR-937-3p55506571
hsa-miR-937-5p55516572
hsa-miR-93855526573
hsa-miR-939-3p55536574
hsa-miR-939-5p55546575
hsa-miR-9-3p55556576
hsa-miR-94055566577
hsa-miR-94155576578
hsa-miR-94255586579
hsa-miR-94355596580
hsa-miR-94455606581
hsa-miR-9555616582
hsa-miR-9-5p55626583
hsa-miR-96-3p55636584
hsa-miR-96-5p55646585
hsa-miR-98-3p55656586
hsa-miR-98-5p55666587
hsa-miR-99a-3p55676588
hsa-miR-99a-5p55686589
hsa-miR-99b-3p55696590
hsa-miR-99b-5p55706591

As shown in Table 10, microRNAs are differentially expressed in different tissues and cells, and often associated with different types of diseases (e.g. cancer cells). The decision of removal or insertion of microRNA binding sites, or any combination, is dependent on microRNA expression patterns and their profilings in cancer cells. In Table 10, “HCC” represents hepatocellular carcinoma, “ALL” stands for acute lymphoblastsic leukemia, “RCC” stands for renal cell carcinoma, “CLL” stands for chrominc lymphocytic leukemia and “MALT” stands for mucosa-associated lymphoid tissue.

TABLE 10
mirs, tissues/cell expression and diseases
BS
mirSEQAssociatedBiological
microRNASEQ IDIDTissues/cellsDiseaseFunction
hsa-let-7a-2-3p25083529Embryonic steminflammatory,tumor
cells, lung, myeloidvarious cancerssuppressor
cells(lung, cervical,
breast, pancreatic,
etc)
hsa-let-7a-3p25093530Embryonic steminflammatory,tumor
cells, lungvarious cancerssuppressor
(lung, cervical,
breast, pancreatic,
etc)
hsa-let-7a-5p25103531Embryonic steminflammatory,tumor
cells, lungvarious cancerssuppressor
(lung, cervical,
breast, pancreatic,
etc)
hsa-let-7b-3p25113532epithelial cells,lung cancer,tumor
endothelial cellscolorectal cancer,angiogenesis
(vascular)cervical cancer,
inflammation and
immune response
after infection
hsa-let-7b-5p25123533epithelial cells,cervical cancer,tumor
endothelial cellsinflammation andangiogenesis
(vascular)immune response
after infection
hsa-let-7c25133534dendritic cellsvarious cacnerstumor
(cervical,suppressor,
pancreatic,apoptosis
lung,
esopphageal, etc)
hsa-let-7d-3p25143535embryonic stemassociated withtumor
cellsvarious cancersuppressor
cells
hsa-let-7d-5p25153536embryonic stemassociated withtumor
cellsvarious cancersuppressor
cells
hsa-let-7e-3p25163537immune cellsvarious cancertumor
cells,suppressor
autoimmunity,
endotoxin
tolerance
hsa-let-7e-5p25173538immune cellsvarious cancertumor
cellssuppressor
hsa-let-7f-1-3p25183539immune cells (Tvarious cancertumor
cells)cellssuppressor
hsa-let-7f-2-3p25193540immune cells (Tvarious cancertumor
cells)cellssuppressor
hsa-let-7f-5p25203541immune cells (TVarious cancertumor
cells)cellssuppressor
hsa-let-7g-3p25213542hematopoietic cells,various cancertumor
adipose, smoothcells (lung, breast,suppressor
muscle cellsetc)
hsa-let-7g-5p25223543hematopoietic cells,various cancertumor
adipose, smoothcells (lung, breast,suppressor
muscle cellsetc)
hsa-let-7i-3p25233544immune cellschronictumor
lymphocytesuppressor
leukimia
hsa-let-7i-5p25243545immune cellschronictumor
lymphocytesuppressor
leukimia
hsa-miR-125253546muscle, heartangiogenesis,
cell
proliferation (myogenesis)
hsa-miR-100-3p25263547hematopoietic cells,gastric cancer,tumor
endothelial cellspancreatic cancerangiogenesis
hsa-miR-100-5p25273548hematopoietic cells,gastric cancer,tumor
endothelial cellspancreatic cancerangiogenesis
hsa-miR-101-3p25283549endothelial cellsvarious cancersangiogenesis
(breast, non-small
cell lung, colon,
gastric,
pancreatic,
bladder, etc);
lupus
erythematosus
hsa-miR-101-5p25293550endothelial cellsvarious cancersangiogenesis
(breast, non-small
cell lung, colon,
gastric,
pancreatic,
bladder, etc);
lupus
erythematosus
hsa-miR-103a-2-5p25303551embryonic stemvarious cancersoncogene, cell
cells, many(endometrial,growth
tissues/cellsneuroblastoma,
colorectal, breast,
liver, etc)
hsa-miR-103a-3p25313552embryonic stemvarious cancersoncogene, cell
cells, many(endometrial,growth
tissues/cellsneuroblastoma,
colorectal, breast,
liver, etc)
hsa-miR-103b25323553Many tissues/cellsvarious cancersoncogene, cell
(endometrial,growth
neuroblastoma,
colorectal, breast,
liver, etc)
hsa-miR-105-3p25333554pancreatic cells
hsa-miR-105-5p25343555pancreatic cells
hsa-miR-106a-3p25353556osteogenic cellsosteocarcoma,cell
other cancersdifferentiation
hsa-miR-106a-5p25363557osteogenic cellsosteocarcoma,cell
other cancersdifferentiation
hsa-miR-106b-3p25373558embryonic stemvarious cancersoncogene
cells(non-small lung
cancer,
gastric cancer,
HCC, gliomas,
etc)
hsa-miR-106b-5p25383559embryonic stemvarious cancersoncogene
cells(non-small lung
cancer,
gastric cancer,
HCC, gliomas,
etc)
hsa-miR-10725393560many tissues, brainbreast cancer,
hepatocytes/liverpituitary
adenoma,
obesity/diabetes
hsa-miR-10a-3p25403561hematopoeitic cellsacute myeoidoncogene, cell
leukemiagrowth
hsa-miR-10a-5p25413562hematopoeitic cellsacute myeoidoncogene, cell
leukemiagrowth
hsa-miR-10b-3p25423563multiple tissues andvarious cancersoncogene
cells(breast, ovarian,
glioblastoma,
pancreatc ductal
adenocarcinoma,
gastric, etc)
hsa-miR-10b-5p25433564multiple tissues andvarious cancersoncogene
cells(breast, ovarian,
glioblastoma,
pancreatc ductal
adenocarcinoma,
gastric, etc)
hsa-miR-1178-3p25443565osteocarcoma
hsa-miR-1178-5p25453566osteocarcoma
hsa-miR-117925463567osteocarcoma
hsa-miR-118025473568discovered in
sarcoma, no
expression data
hsa-miR-118125483569downregulated in
ovarian cancer
cells,
associated with
HCV infection in
hepatocytes
hsa-miR-118225493570placenta
hsa-miR-118325503571associated with
rectal cancer
hsa-miR-118425513572Hematopoietic cellsdownregulated in
oral leukoplakia
(OLK)
hsa-miR-1185-1-3p25523573placenta
hsa-miR-1185-2-3p25533574placenta
hsa-miR-1185-5p25543575placenta
hsa-miR-119325553576melanoma
hsa-miR-119725563577neublastoma
hsa-miR-120025573578chronic
lynphocytic
leukemia
hsa-miR-120225583579chronic
lynphocytic
leukemia,
downregulated in
ovarian cancer
cells
hsa-miR-120325593580in the
chromosome
8q24 region,
cancer cells
hsa-miR-120425603581in the
chromosome
8q24 region,
cancer cells
hsa-miR-120525613582in the
chromosome
8q24 region,
cancer cells
hsa-miR-120625623583in the
chromosome
8q24 region,
cancer cells
hsa-miR-1207-3p25633584in the
chromosome
8q24 region,
cancer cells
hsa-miR-1207-5p25643585in the
chromosome
8q24 region,
cancer cells
hsa-miR-120825653586in the
chromosome
8q24 region,
cancer cells
hsa-miR-122-3p25663587kidney,Renal Celllipid
liver/hepatocytesCarcinomametabolism
(RCC), cancer
cells
hsa-miR-1224-3p25673588Lupus nephritis
hsa-miR-1224-5p25683589rectal cancer
hsa-miR-1225-3p25693590adrenal
pheochromocytomas;
upregulated in
MITF
KnockDown
melanocytes
hsa-miR-1225-5p25703591prostate cancer
hsa-miR-122-5p25713592liver/hepatocytescancer cellslipid
metabolism
hsa-miR-1226-3p25723593discovered in a
mirtron screening
hsa-miR-1226-5p25733594discovered in a
mirtron screening
hsa-miR-1227-3p25743595cartilage/chondrocytes
hsa-miR-1227-5p25753596cartilage/chondrocytes
hsa-miR-1228-3p25763597liver (hepatocytes)Hepatocellularanti-apoptosis
carcinoma (HCC)
hsa-miR-1228-5p25773598liver (hepatocytes)Hepatocellularanti-apoptosis
carcinoma (HCC)
hsa-miR-1229-3p25783599discovered in a
mirtron screening
hsa-miR-1229-5p25793600discovered in a
mirtron screening
hsa-miR-123125803601HCC
hsa-miR-1233-1-5p25813602serum
hsa-miR-1233-3p25823603serum
hsa-miR-1234-3p25833604discovered in
embryonic stem
cell
hsa-miR-1234-5p25843605discovered in
embryonic stem
cell
hsa-miR-1236-3p25853606lymphatictarget to
endothelial cellsVEGFR-3
hsa-miR-1236-5p25863607lymphatictarget to
endothelial cellsVEGFR-3
hsa-miR-1237-3p25873608esophageal cell line
KYSE-150R
hsa-miR-1237-5p25883609esophageal cell line
KYSE-150R
hsa-miR-1238-3p25893610colorectal cancer
hsa-miR-1238-5p25903611colorectal cancer
hsa-miR-124325913612discovered in
embryonic stem
cells
hsa-miR-124-3p25923613brain, plasmagliomacell
(exosomal)differentiation
hsa-miR-124425933614discovered in
embryonic stem
cells
hsa-miR-1245a25943615discovered in
embryonic stem
cells
hsa-miR-1245b-3p25953616discovered in
embryonic stem
cells
hsa-miR-1245b-5p25963617discovered in
embryonic stem
cells
hsa-miR-124-5p25973618brain, Plasmaupregulated incell
(circulating)heart dysfunction,differentiation
glioma
hsa-miR-124625983619embryonic stem
cells, epithelial
cells
hsa-miR-1247-3p25993620embryoid body
cells
hsa-miR-1247-5p26003621embryoid body
cells
hsa-miR-124826013622component of
SnoRNAs
hsa-miR-124926023623liver (hepatocytes)
hsa-miR-125026033624oligodendrocytes
hsa-miR-125126043625discovered in
embryonic stem
cells
hsa-miR-125226053626discovered in
embryonic stem
cells
hsa-miR-125326063627discovered in
embryonic stem
cells
hsa-miR-125426073628embryonic stem
cells
hsa-miR-1255a26083629discovered in
embryonic stem
cells
hsa-miR-1255b-2-3p26093630discovered in
embryonic stem
cells
hsa-miR-1255b-5p26103631discovered in
embryonic stem
cells
hsa-miR-125626113632discovered inprostate cancer
embryonic stem
cells
hsa-miR-125726123633discovered inliposarcoma (soft
embryonic stemtissue sarcoma)
cells
hsa-miR-125826133634discovered inbreast cancer and
embryonic stemlung cancer
cells
hsa-miR-125a-3p26143635brain,various cancercell proliferation
hematopoietic cells(prostate, HCC,and
etc)differentiation
hsa-miR-125a-5p26153636brain,various cancercell proliferation
hematopoietic cells(prostate, HCC,and
etc)differentiation
hsa-miR-125b-1-3p26163637hematopoietic cellsvarious canceroncogene, cell
(monocytes),(prostate, HCC,differentiation
brain (neuron)etc)
hsa-miR-125b-2-3p26173638hematopoietic cellsvarious canceroncogene, cell
(monocytes),(prostate, HCC,differentiation
brain (neuron)etc)
hsa-miR-125b-5p26183639hematopoietic cells,various canceroncogene, cell
brain (neuron)(cutaneous T celldifferentiation
lymphoma,
prostate, HCC,
etc)
hsa-miR-1260a26193640periodontal tissue
hsa-miR-1260b26203641periodontal tissue
hsa-miR-126126213642embryonic stem
cells
hsa-miR-126226223643embryoid body
cells
hsa-miR-126326233644discovered in
embryonic stem
cells
hsa-miR-126-3p26243645endothelialB-lieage ALLangiogenesis
cells, lung
hsa-miR-126426253646discovered in
embryonic stem
cells
hsa-miR-126526263647discovered in
embryonic stem
cells
hsa-miR-126-5p26273648endothelialbreast cancer, B-angiogenesis
cells, lunglieage ALL
hsa-miR-126626283649embryonic stem
cells
hsa-miR-126726293650discovered in
embryonic stem
cells
hsa-miR-1268a26303651embryonic stem
cells
hsa-miR-1268b26313652embryonic stem
cells
hsa-miR-1269a26323653embryoid body
cells
hsa-miR-1269b26333654embryoid body
cells
hsa-miR-127026343655discovered in
embryonic stem
cells
hsa-miR-1271-3p26353656brainHepatocellularSuppress GPC-3
carcinoma (HCC)in HCC
hsa-miR-1271-5p26363657brainHepatocellularSuppress GPC-3
carcinoma (HCC)in HCC
hsa-miR-127226373658embryonic stem
cells
hsa-miR-1273a26383659discovered in
embryonic stem
cells
hsa-miR-1273c26393660colorectal cancer
hsa-miR-1273d26403661discovered in
embryonic stem
cells
hsa-miR-1273e26413662solid tumor cells
hsa-miR-1273f26423663cervical cancer
hsa-miR-1273g-3p26433664cervical cancer
hsa-miR-1273g-5p26443665cervical cancer
hsa-miR-127-3p26453666lung, placenta
hsa-miR-127526463667embryonic stemgastric carcinoma
cells
hsa-miR-127-5p26473668lung, placenta (islet)
hsa-miR-127626483669discovered in
embryonic stem
cells
hsa-miR-1277-3p26493670embryoid body
cells
hsa-miR-1277-5p26503671embryoid body
cells
hsa-miR-127826513672discovered in
embryonic stem
cells
hsa-miR-127926523673monocytes
hsa-miR-12826533674glioblast, brainB-lieage ALLtarget to
neurofibrominlin
neuron
hsa-miR-128126543675muscle invasive
bladder cancer
hsa-miR-128226553676discovered in
embryonic stem
cells
hsa-miR-128326563677placenta
hsa-miR-128426573678lung cancer
hsa-miR-1285-3p26583679various cancerinhibit P53
cellsexpression
hsa-miR-1285-5p26593680various cancerinhibit P53
cellsexpression
hsa-miR-128626603681smooth muscleesophageal cancer
hsa-miR-128726613682embryoid bodybreast cancer
cells
hsa-miR-128826623683discovered in
embryonic stem
cells
hsa-miR-128926633684multiple cell types
hsa-miR-129026643685embryoid bodygastric carcinoma
cells
hsa-miR-129126653686hepatocytescomponent of
SnoRNAs
hsa-miR-129-1-3p26663687multiple cell typesHCC cancer cells
hsa-miR-1292-3p26673688
hsa-miR-129-2-3p26683689multiple cell typesvarious cancer
cells
hsa-miR-1292-5p26693690
hsa-miR-129326703691discovered in
embryonic stem
cells
hsa-miR-129426713692discovered in
embryonic stem
cells
hsa-miR-1295a26723693tumor cells
(follicular
lymphoma)
hsa-miR-1295b-3p26733694tumor cells
(follicular
lymphoma)
hsa-miR-1295b-5p26743695tumor cells
(follicular
lymphoma)
hsa-miR-129-5p26753696liver (hepatocytes)HCC, thyroidcell death in
cancercancer cell
hsa-miR-129626763697breast cancer
hsa-miR-129726773698discovered in
embryonic stem
cells
hsa-miR-129826783699
hsa-miR-129926793700discovered in
embryonic stem
cells
hsa-miR-130126803701breast cancer
hsa-miR-130226813702
hsa-miR-130326823703hepatocytecolorectal cancer,
liver cancer
hsa-miR-1304-3p26833704dental
development
hsa-miR-1304-5p26843705dental
development
hsa-miR-130526853706discovered in
embryonic stem
cells
hsa-miR-1306-3p26863707discovered in
embryonic stem
cells
hsa-miR-1306-5p26873708discovered in
embryonic stem
cells
hsa-miR-1307-3p26883709discovered in
embryonic stem
cells
hsa-miR-1307-5p26893710discovered in
embryonic stem
cells
hsa-miR-130a-3p26903711lung, monocytes,various cancerspro-angiogenic
vascular endothelial(basal cell
cellscarcinoma,
HCC, ovarian,
etc), drug
resistance
hsa-miR-130a-5p26913712lung, monocytes,various cancerspro-angiogenic
vascular endothelial(basal cell
cellscarcinoma,
HCC, ovarian,
etc), drug
resistance
hsa-miR-130b-3p26923713Lung, epidermalvarious cancerscell
cells (keratinocytes)(gastric, rena cellproiferation/senescence
carcinoma)
hsa-miR-130b-5p26933714Lung, epidermalvarious cancerscell
cells (keratinocytes)(gastric, rena cellproiferation/senescence
carcinoma)
hsa-miR-132126943715neuroblastoma
hsa-miR-132226953716neuroblastoma
hsa-miR-132326963717placentaneuroblastoma
hsa-miR-132-3p26973718Brain (neuron),
immune cells
hsa-miR-132426983719neuroblastoma
hsa-miR-132-5p26993720brain (neuron),
immune cells
hsa-miR-133a27003721muscle, heart,heart failure,myogenesis
epithelial cellsesophageal cancer
(lung)
hsa-miR-133b27013722muscle, heart,heart failure,myogenesis
epithelial cellsesophageal cancer
(lung)
hsa-miR-13427023723lung (epithelial)non-samll cell
lung cancer,
pulmonary
embolism
hsa-miR-134327033724breast cancer cells
hsa-miR-135a-3p27043725brain, other tissuesvarious cancertumor
cells (lung, breast,suppressor
colorectal, HCC,
etc)
hsa-miR-135a-5p27053726brain, other tissuesvarious cancertumor
cells (lung, breast,suppressor
colorectal, HCC,
etc)
hsa-miR-135b-3p27063727brain, placenta,various cancers
other tissues(gastric,
mammary, neuroblastomas,
pancreatic, etc)
hsa-miR-135b-5p27073728brain, placenta,various cancers
other tissues(gastric,
mammary, neuroblastomas,
pancreatic, etc)
hsa-miR-136-3p27083729stem cells, placentagliomatumor
suppressor
hsa-miR-136-5p27093730stem cells, placentagliomatumor
suppressor
hsa-miR-13727103731brainvarious cancersinhibiting
(glioblastoma,cancer cell
breast, gastricproliferation and
etc), Alzheimer'smigration
disease
hsa-miR-138-1-3p27113732stem cells,arious cancercell
epidermalcells,proliferation/senescence
cells (keratinocytes)downregulated in
HCC
hsa-miR-138-2-3p27123733stem cellsarious cancer
cells,
downregulated in
HCC
hsa-miR-138-5p27133734stem cellsarious cancer
cells,
downregulated in
HCC
hsa-miR-139-3p27143735hematocytes, brainvarious cancerrepress cancer
cells (colorectal,metastasis
gastric, ovarian)
hsa-miR-139-5p27153736hematocytes, brainvarious cancerrepress cancer
cells (colorectal,metastasis
gastric, ovarian)
hsa-miR-140-3p27163737airway smoothVirus infection,
musclecancers
hsa-miR-140-5p27173738cartilagecsncers
(chondrocytes)
hsa-miR-141-3p27183739Many tissues/cellsvarious cancercell
cells (HCC,differentiation
prostate, kidney,
etc)
hsa-miR-141-5p27193740Many tissues/cellsvarious cancercell
cells (HCC,differentiation
prostate, kidney,
etc)
hsa-miR-142-3p27203741meyloid cells,immune
hematopoiesis,response
APC cells
hsa-miR-142-5p27213742meyloid cells,immune
hematopoiesis,response
APC cells
hsa-miR-143-3p27223743vascular smoothpre-B-cell acute
musclelymphocytic
leukemia, virus
infection
hsa-miR-143-5p27233744vascular smoothvirus infection
muscle, T-cells
hsa-miR-144-3p27243745erythroidvarious cancerscell
(lung, colorectal,differentiation
etc)
hsa-miR-144-5p27253746erythroidvarious cancerscell
(lung, colorectal,differentiation
etc)
hsa-miR-145-3p27263747kidney, cartilage,T-cell lupustumor
vascular smoothsuppressor
muscle
hsa-miR-145-5p27273748kidney, cartilage,T-cell lupustumor
vascular smoothsuppressor
muscle
hsa-miR-146827283749lung cancer
hsa-miR-146927293750tumor
cell (follicular
lymphoma), rectal
cancer
hsa-miR-146a-3p27303751immune cells,various cancers,
hematopoiesisendotoxin
tolerance
hsa-miR-146a-5p27313752immune cells,various cancers,
hematopoiesisendotoxin
tolerance
hsa-miR-146b-3p27323753immune cellsvarious cancers
hsa-miR-146b-5p27333754Embryonic stemvarious cancerstumor invation,
cells(glioma)migration
hsa-miR-147027343755
hsa-miR-147127353756tumor
cell (follicular
lymphoma), rectal
cancer
hsa-miR-147a27363757Macrophageinflammatory
response
hsa-miR-147b27373758Macrophageinflammatory
response
hsa-miR-148a-3p27383759hematopoietic cellsCLL, T-lineage
ALL
hsa-miR-148a-5p27393760hematopoietic cellsCLL, T-lineage
ALL
hsa-miR-148b-3p27403761neuron
hsa-miR-148b-5p27413762neuron
hsa-miR-149-3p27423763heart, brainvarious cancers
(glioma,
colorectal, gastric,
etc)
hsa-miR-149-5p27433764heart, brainvarious cancers
(glioma,
colorectal, gastric,
etc)
hsa-miR-150-3p27443765hematopoietic cellscirculating plasma
(lymphoid)(acute myeloid
leukemia)
hsa-miR-150-5p27453766hematopoietic cellscirculating plasma
(lymphoid)(acute myeloid
leukemia)
hsa-miR-151a-3p27463767neuron, fetal liver
hsa-miR-151a-5p27473768neuron, fetal liver
hsa-miR-151b27483769immune cells (B-
cells)
hsa-miR-15227493770liver
hsa-miR-15327503771brain
hsa-miR-153727513772
hsa-miR-153827523773bloodCancer cells
hsa-miR-153927533774esophageal cell line
KYSE-150R
hsa-miR-154-3p27543775embryonic stem
cells
hsa-miR-154-5p27553776embryonic stem
cells
hsa-miR-155-3p27563777T/B cells,various cancers
monocytes, breast(CLL, B cell
lymphoma,
breast, lung,
ovarian, cervical,
colorectal,
prostate)
hsa-miR-155-5p27573778T/B cells,various cancers
monocytes, breast(CLL, B cell
lymphoma,
breast, lung,
ovarian, cervical,
colorectal,
prostate)
hsa-miR-158727583779identified in B-cells
hsa-miR-15a-3p27593780blood, lymphocyte,cell cycle,
hematopoieticproliferation
tissues (spleen)
hsa-miR-15a-5p27603781blood, lymphocyte,cell cycle,
hematopoieticproliferation
tissues (spleen)
hsa-miR-15b-3p27613782blood, lymphocyte,cell cycle,
hematopoieticproliferation
tissues (spleen)
hsa-miR-15b-5p27623783blood, lymphocyte,cell cycle,
hematopoieticproliferation
tissues (spleen)
hsa-miR-16-1-3p27633784embryonic stem
cells, blood,
hematopoietic
tissues (spleen)
hsa-miR-16-2-3p27643785blood, lymphocyte,
hematopoietic
tissues (spleen)
hsa-miR-16-5p27653786Many tissues, blood
hsa-miR-17-3p27663787embryonic stemtumor
cells, endothelialangiogenesis
cells,
hsa-miR-17-5p27673788endothelial cells,tumor
kidney, breast;angiogenesis
hsa-miR-181a-2-3p27683789glioblast, stem cells
hsa-miR-181a-3p27693790glioblast, myeloid
cells, Embryonic
stem cells
hsa-miR-181a-5p27703791glioblast, myeloid
cells, Embryonic
stem cells
hsa-miR-181b-3p27713792glioblast,cell
Embryonic stemproiferation/senescence
cells, epidermal
(keratinocytes)
hsa-miR-181b-5p27723793glioblast,cell
Embryonic stemproiferation/senescence
cells, epidermal
(keratinocytes)
hsa-miR-181c-3p27733794brain, stemvariou cance cellscell
cells/progenitor(gliobasltoma,differentiation
basal cell
carcinoma,
prostate)
hsa-miR-181c-5p27743795brain, stemvariou cance cellscell
cells/progenitor(gliobasltoma,differentiation
basal cell
carcinoma,
prostate)
hsa-miR-181d27753796glia cells
hsa-miR-182-3p27763797immune cellsautoimmuneimmune
response
hsa-miR-182527773798discovered in a
MiRDeep screening
hsa-miR-182-5p27783799lung, immune cellsautoimmuneimmune
response
hsa-miR-182727793800small cell lung
cancer
hsa-miR-183-3p27803801brain
hsa-miR-183-5p27813802brain
hsa-miR-18427823803blood, tongue,
pancreas (islet)
hsa-miR-185-3p27833804
hsa-miR-185-5p27843805
hsa-miR-186-3p27853806osteoblasts, heartvarious cancer
cells
hsa-miR-186-5p27863807osteoblasts, heartvarious cancer
cells
hsa-miR-187-3p27873808thyroid tumor
hsa-miR-187-5p27883809thyroid tumor
hsa-miR-188-3p27893810irway smooth
muscle, central
nervous system
hsa-miR-188-5p27903811irway smooth
muscle, central
nervous system
hsa-miR-18a-3p27913812endothelial cells,
lung
hsa-miR-18a-5p27923813endothelial cells,
lung
hsa-miR-18b-3p27933814lung
hsa-miR-18b-5p27943815lung
hsa-miR-190827953816breast cancer
hsa-miR-1909-3p27963817rectal cancer
hsa-miR-1909-5p27973818rectal cancer
hsa-miR-190a27983819brain
hsa-miR-190b27993820brain
hsa-miR-191028003821embryonic stem
cells
hsa-miR-1911-3p28013822embryonic stem
cells, neural
precursor
hsa-miR-1911-5p28023823embryonic stem
cells, neural
precursor
hsa-miR-191228033824embryonic stem
cells, neural
precursor
hsa-miR-191328043825embryonic stem
cells
hsa-miR-191-3p28053826chroninc
lymphocyte
leukimia, B-
lieage ALL
hsa-miR-1914-3p28063827embryonic stem
cells
hsa-miR-1914-5p28073828embryonic stem
cells
hsa-miR-1915-3p28083829embryonic stem
cells
hsa-miR-1915-5p28093830embryonic stem
cells
hsa-miR-191-5p28103831chroninc
lymphocyte
leukimia, B-
lieage ALL
hsa-miR-192-3p28113832kidney
hsa-miR-192-5p28123833kidney
hsa-miR-193a-3p28133834many tissues/cellsvarious cancertumor
cells (lung,suppressor,
osteoblastoma,proliferation
ALL, follicular
lymphoma, etc)
hsa-miR-193a-5p28143835many tissues/cellsvarious cancertumor
cells (lung,suppressor,
osteoblastoma,proliferation
ALL, follicular
lymphoma, etc)
hsa-miR-193b-3p28153836many tissues/cells,arious cancertumor
semencells (prostate,suppressor
breast, melanoma,
myeloma, non
small cell lung,
etc)follicular
lymphoma)
hsa-miR-193b-5p28163837many tissues/cells,arious cancertumor
semencells (prostate,suppressor
breast, melanoma,
myeloma, non
small cell lung,
etc)follicular
lymphoma)
hsa-miR-194-3p28173838kidney, livervarious cancers
hsa-miR-194-5p28183839kidney, livervarious cancers
hsa-miR-195-3p28193840breast, pancreas
(islet)
hsa-miR-195-5p28203841breast, pancreas
(islet)
hsa-miR-196a-3p28213842pancreaticvarious canceroncogenic,
cells, endometrialcells (pancreatic,tumor
tissues,osteosarcoma,suppressor
mesenchymal stemendometrial,
cellsAML etc)
hsa-miR-196a-5p28223843pancreaticvarious canceroncogenic,
cells, endometrialcells (pancreatic,tumor
tissues,osteosarcoma,suppressor
mesenchymal stemendometrial,
cellsAML etc)
hsa-miR-196b-3p28233844endometrial tissuesglioblastomaapoptosis
hsa-miR-196b-5p28243845endometrial tissuesglioblastomaapoptosis
hsa-miR-197228253846acute
lymphoblastic
leukemia
hsa-miR-197328263847acute
lymphoblastic
leukemia
hsa-miR-197-3p28273848blood (myeloid),various cancers
other tissues/cells(thyroid tumor,
leukemia, etc)
hsa-miR-197-5p28283849blood (myeloid),various cancers
other tissues/cells(thyroid tumor,
leukemia, etc)
hsa-miR-197628293850acute
lymphoblastic
leukemia
hsa-miR-19828303851central nevous
system (CNS)
hsa-miR-199a-3p28313852liver, embryoid
body cells,
cardiomyocytes
hsa-miR-199a-5p28323853liver,
cardiomyocytes
hsa-miR-199b-3p28333854liver, osteoblastvarious cancersosteogenesis
hsa-miR-199b-5p28343855liver, osteoblastvarious cancersosteogenesis
hsa-miR-19a-3p28353856endothelial cellstumor
angiogenesis
hsa-miR-19a-5p28363857endothelial cellstumor
angiogenesis
hsa-miR-19b-1-5p28373858endothelial cellstumor
angiogenesis
hsa-miR-19b-2-5p28383859endothelial cellstumor
angiogenesis
hsa-miR-19b-3p28393860endothelial cellstumor
angiogenesis
hsa-miR-200a-3p28403861epithelial cells,various cancerstumor
many other tissues(breast, cervical,progression and
bladder, etc)metastasis
hsa-miR-200a-5p28413862epithelial cells,various cancerstumor
many other tissues(breast, cervical,progression and
bladder, etc)metastasis
hsa-miR-200b-3p28423863epithelial cells,tumor
many other tissuesprogression and
metastasis
hsa-miR-200b-5p28433864epithelial cells,tumor
many other tissuesprogression and
metastasis
hsa-miR-200c-3p28443865epithelial cells,tumor
many other tissues,progression and
embryonic stemmetastasis
cells
hsa-miR-200c-5p28453866epithelial cells,tumor
many other tissues,progression and
embryonic stemmetastasis
cells
hsa-miR-202-3p28463867bloodlymphomagenesis,
other cancers
hsa-miR-202-5p28473868bloodlymphomagenesis,
other cancers
hsa-miR-203a28483869skin (epithelium)psoriasis,
autoimmune
hsa-miR-203b-3p28493870skin specificpsoriasis,
(epithelium)autoimmune
hsa-miR-203b-5p28503871skin specificpsoriasis,
(epithelium)autoimmune
hsa-miR-204-3p28513872adipose, othervarious cancerstumor
tissues/cells, kidneymetastasis
hsa-miR-204-5p28523873adipose, othervarious cancerstumor
tissues/cells, kidneymetastasis
hsa-miR-205228533874
hsa-miR-205328543875
hsa-miR-205-3p28553876blood (plasma)various cancer
cells (breast,
glioma,
melanoma,
endometrial, etc)
hsa-miR-205428563877
hsa-miR-205-5p28573878blood (plasma)various cancer
cells (breast,
glioma,
melanoma,
endometrial, etc)
hsa-miR-20628583879muscle (cardiac andmyogenesis
skeletal)
hsa-miR-208a28593880heart (cardiomyocyte),cardiac defects
muscle
hsa-miR-208b28603881heart (cardiomyocyte),cardiac defects
muscle
hsa-miR-20a-3p28613882endothelial cells,
kidney, osteogenic
cells
hsa-miR-20a-5p28623883endothelial cells,
kidney, osteogenic
cells
hsa-miR-20b-3p28633884osteogenic cells
hsa-miR-20b-5p28643885osteogenic cells
hsa-miR-21028653886kidney, heart,RCC, B-cellangiogenesis
vascular endotheliallymphocytes
cells
hsa-miR-211028663887rectal cancer
hsa-miR-211328673888embryonic stem
cells
hsa-miR-211-3p28683889melanocytesmelanoma and
other cancers
hsa-miR-2114-3p28693890ovary, female
reproductuve tract
hsa-miR-2114-5p28703891ovary, female
reproductuve tract
hsa-miR-2115-3p28713892female reproductiveovarian cancer
tract
hsa-miR-2115-5p28723893female reproductiveovarian cancer
tract
hsa-miR-211-5p28733894melanocytesmelanoma and
other cancers
hsa-miR-2116-3p28743895live
cancer (hepatocytes)
and ovarian
cancer
hsa-miR-2116-5p28753896live
cancer (hepatocytes)
and ovarian
cancer
hsa-miR-211728763897ovarian cancer
hsa-miR-212-3p28773898brain (neuron),lymphoma
spleen
hsa-miR-212-5p28783899brain (neuron),lymphoma
spleen
hsa-miR-21-3p28793900glioblast, Bloodautoimmune,
(meyloid cells),heart diseases,
liver, vascularcancers
endothelial cells
hsa-miR-214-3p28803901immune cerlls,varioua cancersimmune
pancreas(melanoma,response
pancreatic,
ovarian)
hsa-miR-214-5p28813902immune cells,varioua cancersimmune
pancreas(melanoma,response
pancreatic,
ovarian)
hsa-miR-21528823903many tissues/cellsvarious cancerscell cycle
(renal, colon,arrest/p53
osteosarcoma)inducible
hsa-miR-21-5p28833904blood (myeloidautoimmune,
cells), liver,heart diseases,
endothelial cellscancers
hsa-miR-216a-3p28843905kidney, pancreas
hsa-miR-216a-5p28853906kidney, pancreas
hsa-miR-216b28863907cancerssenescence
hsa-miR-21728873908endothelial cellsvarious cancer
cells (pancreas,
kidney, breast)
hsa-miR-218-1-3p28883909endothelial cellsvarious cancer
cells (gastric
tumor, bladder,
cervical, etc)
hsa-miR-218-2-3p28893910various cancer
cells (gastric
tumor, bladder,
cervical, etc)
hsa-miR-218-5p28903911various cancer
cells (gastric
tumor, bladder,
cervical, etc)
hsa-miR-219-1-3p28913912brain,
oligodendrocytes
hsa-miR-219-2-3p28923913brain,
oligodendrocytes
hsa-miR-219-5p28933914brain,
oligodendrocytes
hsa-miR-221-3p28943915endothelial cells,leukemia andangiogenesis/vasculogenesis
immune cellsother cancers
hsa-miR-221-5p28953916endothelial cells,leukemia andangiogenesis/vasculogenesis
immune cellsother cancers
hsa-miR-222-3p28963917endothelial cellsvarious cancersangiogenesis
hsa-miR-222-5p28973918endothelial cellsvarious cancersangiogenesis
hsa-miR-223-3p28983919meyloid cellsleukemia
hsa-miR-223-5p28993920meyloid cellsleukemia
hsa-miR-22-3p29003921many tissues/cellsvarious cancerstumorigenesis
hsa-miR-224-3p29013922blood (plasma),cancers and
ovaryinflammation
hsa-miR-224-5p29023923blood (plasma),cancers and
ovaryinflammation
hsa-miR-22-5p29033924many tissues/cellsVarious cancerstumorigenesis
hsa-miR-227629043925breast cancer
hsa-miR-2277-3p29053926female reproductive
tract
hsa-miR-2277-5p29063927female reproductive
tract
hsa-miR-227829073928breast cancer
hsa-miR-2355-3p29083929embryonic stem
cells
hsa-miR-2355-5p29093930embryonic stem
cells
hsa-miR-239229103931identified in B-cells
hsa-miR-23a-3p29113932brain (astrocyte),Cancers
endothelial cells,
blood (erythroid)
hsa-miR-23a-5p29123933brain (astrocyte),cancers
endothelial cells,
blood (erythroid)
hsa-miR-23b-3p29133934blood, meyloidcancers (renal
cellscancer,
glioblastoma,
prostate, etc)
and autoimmune
hsa-miR-23b-5p29143935blood, meyloidcancers (glioblastoma,
cellsprostate, etc)
and autoimmune
hsa-miR-23c29153936cervical cancer
hsa-miR-24-1-5p29163937lung, meyloid cells
hsa-miR-24-2-5p29173938lung, meyloid cells
hsa-miR-24-3p29183939lung, meyloid cells
hsa-miR-2467-3p29193940breast cancer
hsa-miR-2467-5p29203941breast cancer
hsa-miR-25-3p29213942embryonic stem
cells, airway
smooth muscle
hsa-miR-25-5p29223943embryonic stem
cells, airway
smooth muscle
hsa-miR-2681-3p29233944breast cancer
hsa-miR-2681-5p29243945breast cancer
hsa-miR-2682-3p29253946
hsa-miR-2682-5p29263947
hsa-miR-26a-1-3p29273948embryonic stemCLL and othercell cycle and
cells, blood, othercancersdifferentiation
tissues
hsa-miR-26a-2-3p29283949blood, other tissuesCLL and othercell cycle and
cancersdifferentiation
hsa-miR-26a-5p29293950blood, other tissuesCLL and othercell cycle and
cancersdifferentiation
hsa-miR-26b-3p29303951hematopoietic cells
hsa-miR-26b-5p29313952hematopoietic cells
hsa-miR-27a-3p29323953meyloid cellsvarious cancer
cells
hsa-miR-27a-5p29333954meyloid cellsvarious cancer
cells
hsa-miR-27b-3p29343955meyloid cells,various cancerpro-angiogenic
vascular endothelialcells
cells
hsa-miR-27b-5p29353956meyloid cells,various cancerpro-angiogenic
vascular endothelialcells
cells
hsa-miR-28-3p29363957blood (immuneB/T cell
cells)lymphoma
hsa-miR-28-5p29373958blood (immuneB/T cell
cells)lymphoma
hsa-miR-286129383959osteoblastsbasal cell
carcinoma
hsa-miR-290929393960T-Lymphocytes
hsa-miR-296-3p29403961kidney, heart, lung,angiogenesis
entothelial cells
hsa-miR-2964a-3p29413962
hsa-miR-2964a-5p29423963
hsa-miR-296-5p29433964lung, liver,angiogenesis
endothelial cells
hsa-miR-29729443965oocyte and prostate
hsa-miR-29829453966breast cancer
hsa-miR-299-3p29463967myeloid
leukaemia,
hepatoma, breast
cancer
hsa-miR-299-5p29473968myeloid
leukaemia,
hepatoma, breast
cancer
hsa-miR-29a-3p29483969immuno systemCLL, othertumor
cancers,suppression,
neurodegenativeimmune
diseasemodulation
hsa-miR-29a-5p29493970immuno systemCLL, othertumor
cancers,suppression,
neurodegenativeimmune
diseasemodulation
hsa-miR-29b-1-5p29503971immuno systemCLL, othertumor
cancers,suppression,
neurodegenativeimmune
diseasemodulation
hsa-miR-29b-2-5p29513972immuno systemCLL, othertumor
cancerssuppression,
immune
modulation
hsa-miR-29b-3p29523973immuno systemCLL, othertumor
cancerssuppression,
immune
modulation
hsa-miR-29c-3p29533974immuno systemCLL, othertumor
cancerssuppression,
immune
modulation
hsa-miR-29c-5p29543975immuno systemCLL, othertumor
cancerssuppression,
immune
modulation
hsa-miR-30029553976osteoblastBladder cancer
hsa-miR-301a-3p29563977embryonic stem
cells
hsa-miR-301a-5p29573978embryonic stem
cells
hsa-miR-301b29583979esophageal
adenocarcinoma,
colonic cancer
hsa-miR-302a-3p29593980embryonic stemlipid
cells, lipidmetabolism
metabolism
hsa-miR-302a-5p29603981embryonic stemlipid
cells, lipidmetabolism
metabolism
hsa-miR-302b-3p29613982embryonic stem
cells
hsa-miR-302b-5p29623983embryonic stem
cells
hsa-miR-302c-3p29633984embryonic stem
cells
hsa-miR-302c-5p29643985embryonic stem
cells
hsa-miR-302d-3p29653986embryonic stem
cells
hsa-miR-302d-5p29663987embryonic stem
cells
hsa-miR-302e29673988embryoid body
cells
hsa-miR-302f29683989gastric cancer
hsa-miR-3064-3p29693990
hsa-miR-3064-5p29703991
hsa-miR-3065-3p29713992oligodendrocytesanti-virus
response
hsa-miR-3065-5p29723993oligodendrocytessolid tumors
hsa-miR-3074-3p29733994various
cancer (melanoma,
breast)
hsa-miR-3074-5p29743995various
cancer (melanoma,
breast)
hsa-miR-30a-3p29753996kidney, pancreaticvarious cancersautophagy
cells
hsa-miR-30a-5p29763997CNS (prefrontalglioma, colonautophagy
cortex), othercarcinoma
tissues
hsa-miR-30b-3p29773998kidney, adipose,
CNS (prefrontal
cortex)
hsa-miR-30b-5p29783999kidney, adipose,
CNS (prefrontal
cortex)
hsa-miR-30c-1-3p29794000kidney, adipose,
CNS (prefrontal
cortex)
hsa-miR-30c-2-3p29804001kidney, adipose,
CNS (prefrontal
cortex)
hsa-miR-30c-5p29814002kidney, adipose,
CNS (prefrontal
cortex)
hsa-miR-30d-3p29824003CNS (prefrontal
cortex
hsa-miR-30d-5p29834004CNS (prefrontal
cortex, embryoid
body cells
hsa-miR-30e-3p29844005myeloid cells, glia
cells
hsa-miR-30e-5p29854006myeloid cells, glia
cells
hsa-miR-311529864007various cancer
(melanoma,
breast tumor)
hsa-miR-311629874008discovered in the
melanoma
miRNAome
hsa-miR-3117-3p29884009discovered in the
melanoma
miRNAome
hsa-miR-3117-5p29894010discovered in the
melanoma
miRNAome
hsa-miR-311829904011discovered in the
melanoma
miRNAome
hsa-miR-311929914012discovered in the
melanoma
miRNAome
hsa-miR-3120-3p29924013discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3120-5p29934014discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3121-3p29944015discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3121-5p29954016discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-312229964017discovered in the
melanoma
miRNAome
hsa-miR-312329974018discovered in the
melanoma
miRNAome
hsa-miR-3124-3p29984019discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3124-5p29994020discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-312530004021discovered in the
melanoma
miRNAome
hsa-miR-3126-3p30014022discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3126-5p30024023discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3127-3p30034024discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3127-5p30044025discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-312830054026discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3129-3p30064027discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3129-5p30074028discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3130-3p30084029discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3130-5p30094030discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-313130104031discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-313230114032discovered in the
melanoma
miRNAome
hsa-miR-313330124033discovered in the
melanoma
miRNAome
hsa-miR-313430134034discovered in the
melanoma
miRNAome
hsa-miR-3135a30144035discovered in the
melanoma
miRNAome
hsa-miR-3135b30154036discovered in B
cells
hsa-miR-3136-3p30164037discovered in thelymphoblastic
melanomaleukaemia and
miRNAomebreast tumor
hsa-miR-3136-5p30174038discovered in thelymphoblastic
melanomaleukaemia and
miRNAomebreast tumor
hsa-miR-313730184039discovered in the
melanoma
miRNAome
hsa-miR-313830194040discovered in the
melanoma
miRNAome, ovary
hsa-miR-313930204041discovered in the
melanoma
miRNAome
hsa-miR-31-3p30214042
hsa-miR-3140-3p30224043discovered in thelymphoblastic
melanomaleukaemia and
miRNAome, ovarybreast tumor
hsa-miR-3140-5p30234044discovered in thelymphoblastic
melanomaleukaemia and
miRNAome, ovarybreast tumor
hsa-miR-314130244045discovered in the
melanoma
miRNAome
hsa-miR-314230254046discovered in the
melanoma
miRNAome;
immune cells
hsa-miR-314330264047discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3144-3p30274048discovered in the
melanoma
miRNAome, ovary
hsa-miR-3144-5p30284049discovered in the
melanoma
miRNAome, ovary
hsa-miR-3145-3p30294050discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3145-5p30304051discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-314630314052discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-314730324053discovered in the
melanoma
miRNAome
hsa-miR-314830334054discovered in the
melanoma
miRNAome
hsa-miR-314930344055discovered in the
melanoma
miRNAome, ovary
hsa-miR-3150a-3p30354056discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3150a-5p30364057discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3150b-3p30374058discovered in thebreast tumor and
melanomalymphoblastic
miRNAomeleukaemia
hsa-miR-3150b-5p30384059discovered in thebreast tumor and
melanomalymphoblastic
miRNAomeleukaemia
hsa-miR-315130394060discovered in thelymphoblastic
melanomaleukaemia
miRNAome
hsa-miR-3152-3p30404061discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3152-5p30414062discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-315330424063discovered in the
melanoma
miRNAome
hsa-miR-315430434064discovered in thelymphoblastic
melanomaleukaemia
miRNAome
hsa-miR-3155a30444065discovered in the
melanoma
miRNAome
hsa-miR-3155b30454066discovered in B
cells
hsa-miR-3156-3p30464067discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3156-5p30474068discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3157-3p30484069discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3157-5p30494070discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3158-3p30504071discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3158-5p30514072discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-315930524073discovered in the
melanoma
miRNAome
hsa-miR-31-5p30534074various cancer
cells (breast, lung,
prostate)
hsa-miR-3160-3p30544075discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3160-5p30554076discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-316130564077discovered in the
melanoma
miRNAome
hsa-miR-3162-3p30574078discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3162-5p30584079discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-316330594080discovered in the
melanoma
miRNAome
hsa-miR-316430604081discovered in the
melanoma
miRNAome
hsa-miR-316530614082discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-316630624083discovered in the
melanoma
miRNAome
hsa-miR-316730634084discovered in the
melanoma
miRNAome, ovary
hsa-miR-316830644085discovered in the
melanoma
miRNAome
hsa-miR-316930654086discovered in the
melanoma
miRNAome
hsa-miR-317030664087discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-317130674088discovered in the
melanoma
miRNAome, ovary
hsa-miR-3173-3p30684089discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3173-5p30694090discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-317430704091discovered in the
melanoma
miRNAome
hsa-miR-317530714092discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-317630724093discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3177-3p30734094discovered in thebreast tumor and
melanomalymphoblastic
miRNAomeleukaemia
hsa-miR-3177-5p30744095discovered in thebreast tumor and
melanomalymphoblastic
miRNAomeleukaemia
hsa-miR-317830754096discovered in the
melanoma
miRNAome
hsa-miR-317930764097discovered in the
melanoma
miRNAome
hsa-miR-318030774098discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3180-3p30784099discovered in breast
tunor
hsa-miR-3180-5p30794100discovered in breast
tumor
hsa-miR-318130804101discovered in the
melanoma
miRNAome
hsa-miR-318230814102discovered in the
melanoma
miRNAome
hsa-miR-318330824103discovered in the
melanoma
miRNAome
hsa-miR-3184-3p30834104discovered in the
melanoma
miRNAome
hsa-miR-3184-5p30844105discovered in the
melanoma
miRNAome
hsa-miR-318530854106discovered in the
melanoma
miRNAome
hsa-miR-3186-3p30864107discovered in the
melanoma
miRNAome, ovary
hsa-miR-3186-5p30874108discovered in the
melanoma
miRNAome, ovary
hsa-miR-3187-3p30884109discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3187-5p30894110discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-318830904111discovered in the
melanoma
miRNAome
hsa-miR-3189-3p30914112discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3189-5p30924113discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3190-3p30934114discovered in thelymphoblastic
melanomaleukaemia
miRNAome
hsa-miR-3190-5p30944115discovered in thelymphoblastic
melanomaleukaemia
miRNAome
hsa-miR-3191-3p30954116discovered in the
melanoma
miRNAome
hsa-miR-3191-5p30964117discovered in the
melanoma
miRNAome
hsa-miR-319230974118discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-319330984119discovered in the
melanoma
miRNAome
hsa-miR-3194-3p30994120discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-3194-5p31004121discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-319531014122discovered in the
melanoma
miRNAome
hsa-miR-319631024123basal cell
carcinoma
hsa-miR-319731034124discovered in the
melanoma
miRNAome
hsa-miR-319831044125discovered in thebreast tumor
melanoma
miRNAome
hsa-miR-319931054126discovered in the
melanoma
miRNAome
hsa-miR-3200-3p31064127discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-3200-5p31074128discovered in thebreast tumor
melanoma
miRNAome, ovary
hsa-miR-320131084129discovered in the
melanoma
miRNAome,
hsa-miR-320231094130discovered in the
melanoma
miRNAome,epithelial
cell BEAS2B
hsa-miR-320a31104131blood,colon cancer
heart (myocardiac)cells, heart
disease
hsa-miR-320b31114132central nevous
system
hsa-miR-320c31124133chondrocytecartilage
metabolism
hsa-miR-320d31134134cancer stem cells
hsa-miR-320e31144135neural cells
hsa-miR-323a-3p31154136neuronsmyeloid
leukaemia,
mudulla thyroid
carcinoma
hsa-miR-323a-5p31164137neuronsmyeloid
leukaemia,
mudulla thyroid
carcinoma
hsa-miR-323b-3p31174138myeloid
leukaemia
hsa-miR-323b-5p31184139myeloid
leukaemia
hsa-miR-32-3p31194140blood, gliavarious cancers
(lung, kidney,
prostate, etc),
virus infection
hsa-miR-324-3p31204141kidney
hsa-miR-324-5p31214142neuronstumor cells
hsa-miR-32531224143neurons, placenta
hsa-miR-32-5p31234144blood, gliavarious cancers
(lung, kidney,
prostate, etc),
virus infection
hsa-miR-32631244145neuronstumor cells
hsa-miR-32831254146neuron, bloodtumor cells
hsa-miR-32931264147brain and platele
hsa-miR-330-3p31274148various cancers
(prostate,
glioblastoma,
colorectal)
hsa-miR-330-5p31284149various cancers
(prostate,
glioblastoma,
colorectal)
hsa-miR-331-3p31294150gastric cancer
hsa-miR-331-5p31304151lymphocytes
hsa-miR-335-3p31314152kidney, breastRCC, multiple
myeloma
hsa-miR-335-5p31324153kidney, breastRCC, multiple
myeloma
hsa-miR-337-3p31334154lunggastric cancer
hsa-miR-337-5p31344155lung
hsa-miR-338-3p31354156epithelial cells,gastric, rectal
oligodendrocytescancer cells,
osteosarcoma
hsa-miR-338-5p31364157oligodendrocytesgastric cancer
hsa-miR-339-3p31374158immune cell
hsa-miR-339-5p31384159immune cell
hsa-miR-33a-3p31394160pancreatic islet,lipid
lipid metabolismmetabolism
hsa-miR-33a-5p31404161pancreatic islet,lipid
lipid metabolismmetabolism
hsa-miR-33b-3p31414162lipid metabolismlipid
metabolism
hsa-miR-33b-5p31424163lipid metabolismlipid
metabolism
hsa-miR-340-3p31434164various cancers
hsa-miR-340-5p31444165embryoid body
cells
hsa-miR-342-3p31454166brain, circulatingmultiple
plasmamyeloma, other
cancers
hsa-miR-342-5p31464167circulating plasmamultiple
myeloma, other
cancers
hsa-miR-345-3p31474168hematopoietic cellsfollicular
lymphoma, other
cancers
hsa-miR-345-5p31484169hematopoietic cellsfollicular
lymphoma, other
cancers
hsa-miR-34631494170immume cellscancers and
autoimmune
hsa-miR-34a-3p31504171breast, meyloidgastric cancer,tumor
cells, ciliatedCLL, othersuppressor, p53
epithelial cellsinducible
hsa-miR-34a-5p31514172breast, meyloidgastric cancer,tumor
cells, ciliatedCLL, othersuppressor, p53
epithelial cellsinducible
hsa-miR-34b-3p31524173ciliated epithelialvarious cancerstumor
cellssuppressor, p53
inducible
hsa-miR-34b-5p31534174ciliated epithelialvarious cancerstumor
cellssuppressor, p53
inducible
hsa-miR-34c-3p31544175ciliated epithelialvarious cancerstumor
cells, placentasuppressor, p53
inducible
hsa-miR-34c-5p31554176ciliated epithelialvarious cancerstumor
cells, placentasuppressor, p53
inducible
hsa-miR-3529-3p31564177discovered in breast
tumor
hsa-miR-3529-5p31574178discovered in breast
tumor
hsa-miR-3591-3p31584179discovered in breast
tumor
hsa-miR-3591-5p31594180discovered in breast
tumor
hsa-miR-3605-3p31604181discovered in
reprodcutive tracts
hsa-miR-3605-5p31614182discovered in
reprodcutive tracts
hsa-miR-3606-3p31624183discovered in
cervical tumors
hsa-miR-3606-5p31634184discovered in
cervical tumors
hsa-miR-3607-3p31644185discovered in
cervical tumors
hsa-miR-3607-5p31654186discovered in
cervical tumors
hsa-miR-360931664187discovered in
cervical tumors
hsa-miR-361031674188discovered in
cervical tumors
hsa-miR-361131684189discovered in
cervical tumors
hsa-miR-361231694190discovered in
cervical tumors
hsa-miR-3613-3p31704191discovered in
cervical tumors
hsa-miR-3613-5p31714192discovered in
cervical tumors
hsa-miR-361-3p31724193blood, endothelial
cells
hsa-miR-3614-3p31734194discovered in
cervical and breast
tumors
hsa-miR-3614-5p31744195discovered in
cervical and breast
tumors
hsa-miR-361531754196discovered in
cervical tumors
hsa-miR-361-5p31764197endothelial cells
hsa-miR-3616-3p31774198discovered in
cervical tumors
hsa-miR-3616-5p31784199discovered in
cervical tumors
hsa-miR-3617-3p31794200discovered in
cervical tumors and
psoriasis
hsa-miR-3617-5p31804201discovered in
cervical tumors and
psoriasis
hsa-miR-361831814202discovered in
cervical tumors
hsa-miR-3619-3p31824203discovered in breast
tumors
hsa-miR-3619-5p31834204discovered in breast
tumors
hsa-miR-3620-3p31844205discovered in
cervical tumors
hsa-miR-3620-5p31854206discovered in
cervical tumors
hsa-miR-362131864207discovered in
cervical tumors
hsa-miR-3622a-3p31874208discovered in breast
tumors
hsa-miR-3622a-5p31884209discovered in breast
tumors
hsa-miR-3622b-3p31894210discovered in
cervical tumors
hsa-miR-3622b-5p31904211discovered in
cervical tumors
hsa-miR-362-3p31914212melanoma
hsa-miR-362-5p31924213melanoma
hsa-miR-363-3p31934214kidney stem cell,
blood cells
hsa-miR-363-5p31944215kidney stem cell,
blood cells
hsa-miR-364631954216discovered in solid
tumor
hsa-miR-364831964217discovered in solid
tumor
hsa-miR-364931974218discovered in solid
tumor
hsa-miR-365031984219discovered in solid
tumor
hsa-miR-365131994220discovered in solid
tumor
hsa-miR-365232004221discovered in solid
tumor
hsa-miR-365332014222discovered in solid
tumor
hsa-miR-365432024223discovered in solid
tumor
hsa-miR-365532034224discovered in solid
tumor
hsa-miR-365632044225discovered in solid
tumor
hsa-miR-365732054226discovered in solid
tumor
hsa-miR-365832064227discovered in solid
tumor
hsa-miR-365932074228discovered in breast
tumors
hsa-miR-365a-3p32084229various cancerapoptosis
cells (Immune
cells, lung, colon,
endometriotic)
hsa-miR-365a-5p32094230various cancerapoptosis
cells (Immune
cells, lung, colon,
endometriotic))
hsa-miR-365b-3p32104231various cancerapoptosis
(retinoblastoma, colon,
endometriotic)
hsa-miR-365b-5p32114232various cancerapoptosis
(colon,
endometriotic)
hsa-miR-366032124233discovered in breast
tumors
hsa-miR-366132134234discovered in breast
tumors
hsa-miR-366232144235
hsa-miR-3663-3p32154236
hsa-miR-3663-5p32164237
hsa-miR-3664-3p32174238discovered in breast
tumors
hsa-miR-3664-5p32184239discovered in breast
tumors
hsa-miR-366532194240brain
hsa-miR-366632204241brain
hsa-miR-3667-3p32214242discovered in
peripheral blood
hsa-miR-3667-5p32224243discovered in
peripheral blood
hsa-miR-366832234244discovered in
peripheral blood
hsa-miR-366932244245discovered in
peripheral blood
hsa-miR-367032254246discovered in
peripheral blood
hsa-miR-367132264247discovered in
peripheral blood
hsa-miR-367232274248discovered in
peripheral blood
hsa-miR-367332284249discovered in
peripheral blood
hsa-miR-367-3p32294250embryonic stemreprogramming
cells
hsa-miR-367432304251discovered in
peripheral blood
hsa-miR-3675-3p32314252discovered in
peripheral blood
hsa-miR-3675-5p32324253discovered in
peripheral blood
hsa-miR-367-5p32334254embryonic stemreprogramming
cells
hsa-miR-3676-3p32344255discovered in
peripheral blood
hsa-miR-3676-5p32354256discovered in
peripheral blood
hsa-miR-3677-3p32364257discovered in
peripheral blood
hsa-miR-3677-5p32374258discovered in
peripheral blood
hsa-miR-3678-3p32384259discovered in
peripheral blood
hsa-miR-3678-5p32394260discovered in
peripheral blood
hsa-miR-3679-3p32404261discovered in
peripheral blood
hsa-miR-3679-5p32414262discovered in
peripheral blood
hsa-miR-3680-3p32424263discovered in
peripheral blood
hsa-miR-3680-5p32434264discovered in
peripheral blood
hsa-miR-3681-3p32444265discovered in
peripheral blood
hsa-miR-3681-5p32454266discovered in
peripheral blood
hsa-miR-3682-3p32464267discovered in
peripheral blood
hsa-miR-3682-5p32474268discovered in
peripheral blood
hsa-miR-368332484269discovered in
peripheral blood
hsa-miR-368432494270discovered in
peripheral blood
hsa-miR-368532504271discovered in
peripheral blood
hsa-miR-368632514272discovered in
peripheral blood
hsa-miR-368732524273discovered in
peripheral blood
hsa-miR-3688-3p32534274discovered in breast
tumor
hsa-miR-3688-5p32544275discovered in breast
tumor
hsa-miR-3689a-3p32554276discovered in
female
reproductive tract
hsa-miR-3689a-5p32564277discovered in
female
reproductive tract
and peripheral
blood
hsa-miR-3689b-3p32574278discovered in
female
reproductive tract
and peripheral
blood
hsa-miR-3689b-5p32584279discovered in
female
reproductive tract
hsa-miR-3689c32594280discovered in B
cells
hsa-miR-3689d32604281discovered in B
cells
hsa-miR-3689e32614282discovered in B
cells
hsa-miR-3689f32624283discovered in B
cells
hsa-miR-369032634284discovered in
peripheral blood
hsa-miR-3691-3p32644285discovered in
peripheral blood
hsa-miR-3691-5p32654286discovered in
peripheral blood
hsa-miR-3692-3p32664287discovered in
peripheral blood
hsa-miR-3692-5p32674288discovered in
peripheral blood
hsa-miR-369-3p32684289stem cellsreprogramming
hsa-miR-369-5p32694290stem cellsreprogramming
hsa-miR-37032704291acute meyloidtumor
leukaemia andsuppressor, lipid
other cancersmetabolism
hsa-miR-371332714292discovered in
neuroblastoma
hsa-miR-371432724293discovered in
neuroblastoma
hsa-miR-371a-3p32734294serum
hsa-miR-371a-5p32744295serum
hsa-miR-371b-3p32754296serum
hsa-miR-371b-5p32764297serum
hsa-miR-37232774298hematopoietic cells,
lung, placental
(blood)
hsa-miR-373-3p32784299breast cancer
hsa-miR-373-5p32794300breast cancer
hsa-miR-374a-3p32804301muscle (myoblasts)breast and lungmyogenic
cancerdifferentiation
hsa-miR-374a-5p32814302muscle (myoblasts)breast and lungmyogenic
cancerdifferentiation
hsa-miR-374b-3p32824303muscle (myoblasts)myogenic
differentiation
hsa-miR-374b-5p32834304muscle (myoblasts)myogenic
differentiation
hsa-miR-374c-3p32844305muscle (myoblasts)myogenic
differentiation
hsa-miR-374c-5p32854306muscle (myoblasts)myogenic
differentiation
hsa-miR-37532864307pancreas (islet)
hsa-miR-376a-2-5p32874308regulatory miRs for
hematopoietic cells
(erythroid, platelet,
lympho)
hsa-miR-376a-3p32884309regulatory miRs for
hematopoietic cells
(erythroid, platelet,
lympho)
hsa-miR-376a-5p32894310regulatory miRs for
hematopoietic cells
(erythroid, platelet,
lympho)
hsa-miR-376b-3p32904311bloodvarious cancerautophagy
cells
hsa-miR-376b-5p32914312bloodvarious cancerautophagy
cells
hsa-miR-376c-3p32924313trophoblastvarious cancercell proliferatio
cells
hsa-miR-376c-5p32934314trophoblastvarious cancercell proliferatio
cells
hsa-miR-377-3p32944315hematopoietic cells
hsa-miR-377-5p32954316hematopoietic cells
hsa-miR-378a-3p32964317ovary, lipid
metabolism
hsa-miR-378a-5p32974318ovary,
placenta/trophoblast,
lipid metabolism
hsa-miR-378b32984319lipid metabolism
hsa-miR-378c32994320lipid metabolism
hsa-miR-378d33004321lipid metabolism
hsa-miR-378e33014322lipid metabolism
hsa-miR-378f33024323lipid metabolism
hsa-miR-378g33034324lipid metabolism
hsa-miR-378h33044325lipid metabolism
hsa-miR-378i33054326lipid metabolism
hsa-miR-378j33064327lipid metabolism
hsa-miR-379-3p33074328various cancers
(breast,
hepatocytes,
colon)
hsa-miR-379-5p33084329various cancers
(breast,
hepatocytes,
colon)
hsa-miR-380-3p33094330brainneuroblastoma
hsa-miR-380-5p33104331brain, embryonicneuroblastoma
stem cells
hsa-miR-381-3p33114332chondrogenesis,
lung, brain
hsa-miR-381-5p33124333chondrogenesis,
lung, brain
hsa-miR-382-3p33134334renal epithelial cells
hsa-miR-382-5p33144335renal epithelial cells
hsa-miR-38333154336testes, brain
(medulla)
hsa-miR-38433164337epithelial cells
hsa-miR-390733174338discovered in
female reproductive
tract
hsa-miR-390833184339discovered in
female reproductive
tract
hsa-miR-390933194340discovered in
female reproductive
tract
hsa-miR-391033204341discovered in
female reproductive
tract
hsa-miR-391133214342discovered in breast
tumor and female
reproductive tract
hsa-miR-391233224343discovered in
female reproductive
tract
hsa-miR-3913-3p33234344discovered in breast
tumor and female
reproductive tract
hsa-miR-3913-5p33244345discovered in breast
tumor and female
reproductive tract
hsa-miR-391433254346discovered in breast
tumor and female
reproductive tract
hsa-miR-391533264347discovered in
female reproductive
tract
hsa-miR-391633274348discovered in
female reproductive
tract
hsa-miR-391733284349discovered in
female reproductive
tract
hsa-miR-391833294350discovered in
female reproductive
tract
hsa-miR-391933304351discovered in
female reproductive
tract
hsa-miR-392033314352discovered in
female reproductive
tract
hsa-miR-392133324353discovered in
female reproductive
tract
hsa-miR-3922-3p33334354discovered in breast
tumor and female
reproductive tract
hsa-miR-3922-5p33344355discovered in breast
tumor and female
reproductive tract
hsa-miR-392333354356discovered in
female reproductive
tract
hsa-miR-392433364357discovered in
female reproductive
tract
hsa-miR-3925-3p33374358discovered in breast
tumor and female
reproductive tract
hsa-miR-3925-5p33384359discovered in breast
tumor and female
reproductive tract
hsa-miR-392633394360discovered in
female reproductive
tract
hsa-miR-3927-3p33404361discovered in
female reproductive
tract and psoriasis
hsa-miR-3927-5p33414362discovered in
female reproductive
tract and psoriasis
hsa-miR-392833424363discovered in
female reproductive
tract
hsa-miR-392933434364discovered in
female reproductive
tract
hsa-miR-3934-3p33444365discovered in
abnormal skin
(psoriasis)
hsa-miR-3934-5p33454366discovered in
abnormal skin
(psoriasis)
hsa-miR-393533464367
hsa-miR-393633474368discovered in breast
tumor and
lymphoblastic
leukaemia
hsa-miR-393733484369
hsa-miR-393833494370
hsa-miR-393933504371
hsa-miR-3940-3p33514372discovered in breast
tumor
hsa-miR-3940-5p33524373discovered in breast
tumor
hsa-miR-394133534374
hsa-miR-3942-3p33544375discovered in breast
tumor and
lymphoblastic
leukaemia
hsa-miR-3942-5p33554376discovered in breast
tumor and
lymphoblastic
leukaemia
hsa-miR-394333564377
hsa-miR-3944-3p33574378discovered in breast
tumor
hsa-miR-3944-5p33584379discovered in breast
tumor
hsa-miR-394533594380
hsa-miR-396033604381osteoblast
hsa-miR-397233614382discovered in Acute
Myeloid Leukaemia
hsa-miR-397333624383discovered in Acute
Myeloid Leukaemia
hsa-miR-397433634384discovered in Acute
Myeloid Leukaemia
hsa-miR-397533644385discovered in Acute
Myeloid Leukaemia
hsa-miR-397633654386discovered in Acute
Myeloid Leukaemia
hsa-miR-397733664387discovered in Acute
Myeloid Leukaemia
hsa-miR-397833674388discovered in Acute
Myeloid Leukaemia
hsa-miR-409-3p33684389gastric cancer
hsa-miR-409-5p33694390gastric cancer
hsa-miR-41033704391brainglioma
hsa-miR-411-3p33714392Glioblastoma
others
hsa-miR-411-5p33724393Glioblastoma
others
hsa-miR-41233734394upregulated in
lung cancer
hsa-miR-42133744395endothelial cellsgastric cancer,
HCC
hsa-miR-422a33754396circulating
microRNA (in
plasma)
hsa-miR-423-3p33764397embryonic stem
cells
hsa-miR-423-5p33774398heart, embryonic
stem cells
hsa-miR-424-3p33784399endothelial cellsvariouspro-angiogenic
cancers (e.g B-
lieage ALL),
cardiac diseases
hsa-miR-424-5p33794400endothelial cellsvariouspro-angiogenic
cancers (e.g B-
lieage ALL),
cardiac diseases
hsa-miR-425133804401discovered in
embryonic stem
cells and neural
precusors
hsa-miR-425233814402discovered in
embryonic stem
cells and neural
precusors
hsa-miR-425333824403discovered in
embryonic stem
cells and neural
precusors
hsa-miR-425-3p33834404brainovarian cancer,
brain tumor
hsa-miR-425433844405discovered in
embryonic stem
cells and neural
precusors
hsa-miR-425533854406discovered in
embryonic stem
cells and neural
precusors
hsa-miR-425-5p33864407brainB-lieage ALL,
brain tumor
hsa-miR-425633874408discovered in
embryonic stem
cells and neural
precusors
hsa-miR-425733884409discovered in
embryonic stem
cells and neural
precusors
hsa-miR-425833894410discovered in
embryonic stem
cells and neural
precusors
hsa-miR-425933904411discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426033914412discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426133924413discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426233934414discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426333944415discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426433954416discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426533964417discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426633974418discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426733984419discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426833994420discovered in
embryonic stem
cells and neural
precusors
hsa-miR-426934004421discovered in
embryonic stem
cells and neural
precusors
hsa-miR-427034014422discovered in
embryonic stem
cells and neural
precusors
hsa-miR-427134024423discovered in
embryonic stem
cells and neural
precusors
hsa-miR-427234034424discovered in
embryonic stem
cells and neural
precusors
hsa-miR-427334044425
hsa-miR-427434054426discovered in
embryonic stem
cells and neural
precusors
hsa-miR-427534064427discovered in
embryonic stem
cells and neural
precusors
hsa-miR-427634074428discovered in
embryonic stem
cells and neural
precusors
hsa-miR-427734084429discovered in
embryonic stem
cells and neural
precusors
hsa-miR-427834094430discovered in
embryonic stem
cells and neural
precusors
hsa-miR-427934104431discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428034114432discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428134124433discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428234134434discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428334144435discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428434154436discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428534164437discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428634174438discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428734184439discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428834194440discovered in
embryonic stem
cells and neural
precusors
hsa-miR-428934204441discovered in
embryonic stem
cells and neural
precusors
hsa-miR-42934214442Epithelial cellsvarious cancers
(colorectal,
endometrial,
gastric, ovarian
etc)
hsa-miR-429034224443discovered in
embryonic stem
cells and neural
precusors
hsa-miR-429134234444discovered in
embryonic stem
cells and neural
precusors
hsa-miR-429234244445discovered in
embryonic stem
cells and neural
precusors
hsa-miR-429334254446discovered in
embryonic stem
cells and neural
precusors
hsa-miR-429434264447discovered in
embryonic stem
cells and neural
precusors
hsa-miR-429534274448discovered in
embryonic stem
cells and neural
precusors
hsa-miR-429634284449discovered in
embryonic stem
cells and neural
precusors
hsa-miR-429734294450discovered in
embryonic stem
cells and neural
precusors
hsa-miR-429834304451discovered in
embryonic stem
cells and neural
precusors
hsa-miR-429934314452discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430034324453discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430134334454discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430234344455discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430334354456discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430434364457discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430534374458discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430634384459discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430734394460discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430834404461discovered in
embryonic stem
cells and neural
precusors
hsa-miR-430934414462discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431034424463discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431134434464discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431234444465discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431334454466discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431-3p34464467Cancers
(follicular
lymphoma)
hsa-miR-431434474468discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431534484469discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431-5p34494470Cancers
(follicular
lymphoma)
hsa-miR-431634504471discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431734514472discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431834524473discovered in
embryonic stem
cells and neural
precusors
hsa-miR-431934534474discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432034544475discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432134554476discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432234564477discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432334574478discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432-3p34584479myoblastmyogenic
differentiation
hsa-miR-432434594480discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432534604481discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432-5p34614482myoblastmyogenic
differentiation
hsa-miR-432634624483discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432734634484discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432834644485discovered in
embryonic stem
cells and neural
precusors
hsa-miR-432934654486discovered in
embryonic stem
cells and neural
precusors
hsa-miR-43334664487various diseases
(cancer,
Parkinson's,
Chondrodysplasia)
hsa-miR-433034674488discovered in
embryonic stem
cells and neural
precusors
hsa-miR-441734684489discovered in B
cells
hsa-miR-441834694490discovered in B
cells
hsa-miR-4419a34704491discovered in B
cells
hsa-miR-4419b34714492discovered in B
cells
hsa-miR-442034724493discovered in B
cells
hsa-miR-442134734494discovered in B
cells
hsa-miR-442234744495discovered in breast
tumor and B cells
hsa-miR-4423-3p34754496discovered in breast
tumor, B cells and
skin (psoriasis)
hsa-miR-4423-5p34764497discovered in breast
tumor B cells and
skin (psoriasis)
hsa-miR-442434774498discovered in B
cells
hsa-miR-442534784499discovered in B
cells
hsa-miR-442634794500discovered in B
cells
hsa-miR-442734804501discovered in B
cells
hsa-miR-442834814502discovered in B
cells
hsa-miR-442934824503discovered in B
cells
hsa-miR-443034834504discovered in B
cells
hsa-miR-443134844505discovered in B
cells
hsa-miR-443234854506discovered in B
cells
hsa-miR-4433-3p34864507discovered in B
cells
hsa-miR-4433-5p34874508discovered in B
cells
hsa-miR-443434884509discovered in B
cells
hsa-miR-443534894510discovered in B
cells
hsa-miR-4436a34904511discovered in breast
tumor and B cells
hsa-miR-4436b-3p34914512discovered in breast
tumor
hsa-miR-4436b-5p34924513discovered in breast
tumor
hsa-miR-443734934514discovered in B
cells
hsa-miR-443834944515discovered in B
cells
hsa-miR-443934954516discovered in B
cells
hsa-miR-444034964517discovered in B
cells
hsa-miR-444134974518discovered in B
cells
hsa-miR-444234984519discovered in B
cells
hsa-miR-444334994520discovered in B
cells
hsa-miR-444435004521discovered in B
cells
hsa-miR-4445-3p35014522discovered in B
cells
hsa-miR-4445-5p35024523discovered in B
cells
hsa-miR-4446-3p35034524discovered in breast
tumor and B cells
hsa-miR-4446-5p35044525discovered in breast
tumor and B cells
hsa-miR-444735054526discovered in B
cells
hsa-miR-444835064527discovered in B
cells
hsa-miR-444935074528discovered in B
cells
hsa-miR-445035084529discovered in B
cells
hsa-miR-445135094530discovered in B
cells
hsa-miR-445235104531discovered in B
cells
hsa-miR-445335114532discovered in B
cells
hsa-miR-445435124533discovered in B
cells
hsa-miR-445535134534discovered in B
cells
hsa-miR-445635144535discovered in B
cells
hsa-miR-445735154536discovered in B
cells
hsa-miR-445835164537discovered in B
cells
hsa-miR-445935174538discovered in B
cells
hsa-miR-446035184539discovered in B
cells
hsa-miR-446135194540discovered in B
cells
hsa-miR-446235204541discovered in B
cells
hsa-miR-446335214542discovered in B
cells
hsa-miR-446435224543discovered in B
cells
hsa-miR-446535234544discovered in B
cells
hsa-miR-446635244545discovered in B
cells
hsa-miR-446735254546discovered in breast
tumor and B cells
hsa-miR-446835264547discovered in B
cells
hsa-miR-446935274548discovered in breast
tumor and B cells
hsa-miR-447035284549discovered in B
cells
hsa-miR-447145505571discovered in breast
tumor and B cells
hsa-miR-447245515572discovered in B
cells
hsa-miR-447345525573discovered in B
cells
hsa-miR-4474-3p45535574discovered in breast
tumor,
lymphoblastic
leukaemia and B
cells
hsa-miR-4474-5p45545575discovered in breast
tumor,
lymphoblastic
leukaemia and B
cells
hsa-miR-447545555576discovered in B
cells
hsa-miR-447645565577discovered in B
cells
hsa-miR-4477a45575578discovered in B
cells
hsa-miR-4477b45585579discovered in B
cells
hsa-miR-447845595580discovered in B
cells
hsa-miR-447945605581discovered in B
cells
hsa-miR-44845615582liver (hepatocytes)HCC
hsa-miR-448045625583discovered in B
cells
hsa-miR-448145635584discovered in B
cells
hsa-miR-4482-3p45645585discovered in B
cells
hsa-miR-4482-5p45655586discovered in B
cells
hsa-miR-448345665587discovered in B
cells
hsa-miR-448445675588discovered in B
cells
hsa-miR-448545685589discovered in B
cells
hsa-miR-448645695590discovered in B
cells
hsa-miR-448745705591discovered in B
cells
hsa-miR-448845715592discovered in B
cells
hsa-miR-448945725593discovered in breast
tumor and B cells
hsa-miR-449045735594discovered in B
cells
hsa-miR-449145745595discovered in B
cells
hsa-miR-449245755596discovered in B
cells
hsa-miR-449345765597discovered in B
cells
hsa-miR-449445775598discovered in B
cells
hsa-miR-449545785599discovered in B
cells
hsa-miR-449645795600discovered in B
cells
hsa-miR-449745805601discovered in B
cells
hsa-miR-449845815602discovered in B
cells
hsa-miR-449945825603discovered in B
cells
hsa-miR-449a45835604chondrocytes, ciliatedlung, colonic,cell cycle
epithelial cellsovarian cancerprogression and
proliferation
hsa-miR-449b-3p45845605ciliated epithelialvarious cancercell cycle
cells, other tissuescellsprogression and
proliferation
hsa-miR-449b-5p45855606ciliated epithelialvarious cancercell cycle
cells, other tissuescellsprogression and
proliferation
hsa-miR-449c-3p45865607epithelial ovarian
cancer cells
hsa-miR-449c-5p45875608epithelial ovarian
cancer cells
hsa-miR-450045885609discovered in B
cells
hsa-miR-450145895610discovered in B
cells
hsa-miR-450245905611discovered in B
cells
hsa-miR-450345915612discovered in B
cells
hsa-miR-450445925613discovered in B
cells
hsa-miR-450545935614discovered in B
cells
hsa-miR-450645945615discovered in B
cells
hsa-miR-450745955616discovered in B
cells
hsa-miR-450845965617discovered in B
cells
hsa-miR-450945975618discovered in B
cells
hsa-miR-450a-3p45985619
hsa-miR-450a-5p45995620
hsa-miR-450b-3p46005621
hsa-miR-450b-5p46015622
hsa-miR-451046025623discovered in B
cells
hsa-miR-451146035624discovered in B
cells
hsa-miR-451246045625discovered in B
cells
hsa-miR-451346055626discovered in B
cells
hsa-miR-451446065627discovered in B
cells
hsa-miR-451546075628discovered in B
cells
hsa-miR-451646085629discovered in B
cells
hsa-miR-451746095630discovered in B
cells
hsa-miR-451846105631discovered in B
cells
hsa-miR-451946115632discovered in B
cells
hsa-miR-451a46125633heart, central
nevous system,
epithelial cells
hsa-miR-451b46135634heart, central
nevous system,
epithelial cells
hsa-miR-4520a-3p46145635discovered in breast
tumor and B cells,
skin (psoriasis)
hsa-miR-4520a-5p46155636discovered in breast
tumor and B cells,
skin (psoriasis)
hsa-miR-4520b-3p46165637discovered in breast
tumor
hsa-miR-4520b-5p46175638discovered in breast
tumor
hsa-miR-452146185639discovered in B
cells
hsa-miR-452246195640discovered in B
cells
hsa-miR-452346205641discovered in B
cells
hsa-miR-452-3p46215642myoblastbladder cancer
and others
hsa-miR-4524a-3p46225643discovered in breast
tumor and B cells,
skin (psoriasis)
hsa-miR-4524a-5p46235644discovered in breast
tumor and B cells,
skin (psoriasis)
hsa-miR-4524b-3p46245645discovered in breast
tumor and B cells,
skin (psoriasis)
hsa-miR-4524b-5p46255646discovered in breast
tumor and B cells,
skin (psoriasis)
hsa-miR-452546265647discovered in B
cells
hsa-miR-452-5p46275648myoblastbladder cancer
and others
hsa-miR-452646285649discovered in breast
tumor and B cells
hsa-miR-452746295650discovered in B
cells
hsa-miR-452846305651discovered in B
cells
hsa-miR-4529-3p46315652discovered in breast
tumor and B cells
hsa-miR-4529-5p46325653discovered in breast
tumor and B cells
hsa-miR-453046335654discovered in B
cells
hsa-miR-453146345655discovered in B
cells
hsa-miR-453246355656discovered in B
cells
hsa-miR-453346365657discovered in B
cells
hsa-miR-453446375658discovered in B
cells
hsa-miR-453546385659discovered in B
cells
hsa-miR-4536-3p46395660discovered in B
cells
hsa-miR-4536-5p46405661discovered in B
cells
hsa-miR-453746415662discovered in B
cells
hsa-miR-453846425663discovered in B
cells
hsa-miR-453946435664discovered in B
cells
hsa-miR-454046445665discovered in B
cells
hsa-miR-454-3p46455666embryoid body
cells, central
nevous system,
monocytes
hsa-miR-454-5p46465667embryoid body
cells, central
nevous system,
monocytes
hsa-miR-455-3p46475668basal cell
carcinoma, other
cancers
hsa-miR-455-5p46485669basal cell
carcinoma, other
cancers
hsa-miR-4632-3p46495670discovred in breast
tumor
hsa-miR-4632-5p46505671discovered in breast
tumor
hsa-miR-4633-3p46515672discovered in breast
tumor
hsa-miR-4633-5p46525673discovered in breast
tumor
hsa-miR-463446535674discovered in breast
tumor
hsa-miR-463546545675discovered in breast
tumor
hsa-miR-463646555676discovered in breast
tumor
hsa-miR-463746565677discovered in breast
tumor and
lymphoblastic
leukaemia
hsa-miR-4638-3p46575678discovered in breast
tumor
hsa-miR-4638-5p46585679discovered in breast
tumor
hsa-miR-4639-3p46595680discovered in breast
tumor
hsa-miR-4639-5p46605681discovered in breast
tumor
hsa-miR-4640-3p46615682discovered in breast
tumor
hsa-miR-4640-5p46625683discovered in breast
tumor
hsa-miR-464146635684discovered in breast
tumor
hsa-miR-464246645685discovered in breast
tumor
hsa-miR-464346655686discovered in breast
tumor
hsa-miR-464446665687discovered in breast
tumor
hsa-miR-4645-3p46675688discovered in breast
tumor
hsa-miR-4645-5p46685689discovered in breast
tumor
hsa-miR-4646-3p46695690discovered in breast
tumor
hsa-miR-4646-5p46705691discovered in breast
tumor
hsa-miR-464746715692discovered in breast
tumor
hsa-miR-464846725693discovered in breast
tumor
hsa-miR-4649-3p46735694discovered in breast
tumor
hsa-miR-4649-5p46745695discovered in breast
tumor
hsa-miR-4650-3p46755696discovered in breast
tumor
hsa-miR-4650-5p46765697discovered in breast
tumor
hsa-miR-465146775698discovered in breast
tumor
hsa-miR-4652-3p46785699discovered in breast
tumor
hsa-miR-4652-5p46795700discovered in breast
tumor
hsa-miR-4653-3p46805701discovered in breast
tumor
hsa-miR-4653-5p46815702discovered in breast
tumor
hsa-miR-465446825703discovered in breast
tumor
hsa-miR-4655-3p46835704discovered in breast
tumor
hsa-miR-4655-5p46845705discovered in breast
tumor
hsa-miR-465646855706discovered in breast
tumor
hsa-miR-465746865707discovered in breast
tumor
hsa-miR-465846875708discovered in breast
tumor
hsa-miR-4659a-3p46885709discovered in breast
tumor
hsa-miR-4659a-5p46895710discovered in breast
tumor
hsa-miR-4659b-3p46905711discovered in breast
tumor
hsa-miR-4659b-5p46915712discovered in breast
tumor
hsa-miR-46646925713
hsa-miR-466046935714discovered in breast
tumor
hsa-miR-4661-3p46945715discovered in breast
tumor
hsa-miR-4661-5p46955716discovered in breast
tumor
hsa-miR-4662a-3p46965717discovered in breast
tumor, psoriasis
hsa-miR-4662a-5p46975718discovered in breast
tumor, psoriasis
hsa-miR-4662b46985719discovered in breast
tumor
hsa-miR-466346995720discovered in breast
tumor
hsa-miR-4664-3p47005721discovered in breast
tumor
hsa-miR-4664-5p47015722discovered in breast
tumor
hsa-miR-4665-3p47025723discovered in breast
tumor
hsa-miR-4665-5p47035724discovered in breast
tumor
hsa-miR-4666a-3p47045725discovered in breast
tumor
hsa-miR-4666a-5p47055726discovered in breast
tumor
hsa-miR-4666b47065727
hsa-miR-4667-3p47075728discovered in breast
tumor
hsa-miR-4667-5p47085729discovered in breast
tumor
hsa-miR-4668-3p47095730discovered in breast
tumor
hsa-miR-4668-5p47105731discovered in breast
tumor
hsa-miR-466947115732discovered in breast
tumor
hsa-miR-4670-3p47125733discovered in breast
tumor
hsa-miR-4670-5p47135734discovered in breast
tumor
hsa-miR-4671-3p47145735discovered in breast
tumor
hsa-miR-4671-5p47155736discovered in breast
tumor
hsa-miR-467247165737discovered in breast
tumor
hsa-miR-467347175738discovered in breast
tumor
hsa-miR-467447185739discovered in breast
tumor
hsa-miR-467547195740discovered in breast
tumor
hsa-miR-4676-3p47205741discovered in breast
tumor
hsa-miR-4676-5p47215742discovered in breast
tumor
hsa-miR-4677-3p47225743discovered in breast
tumor, psoriasis
hsa-miR-4677-5p47235744discovered in breast
tumor, psoriasis
hsa-miR-467847245745discovered in breast
tumor
hsa-miR-467947255746discovered in breast
tumor
hsa-miR-4680-3p47265747discovered in breast
tumor
hsa-miR-4680-5p47275748discovered in breast
tumor
hsa-miR-468147285749discovered in breast
tumor
hsa-miR-468247295750discovered in breast
tumor
hsa-miR-468347305751discovered in breast
tumor
hsa-miR-4684-3p47315752discovered in breast
tumor
hsa-miR-4684-5p47325753discovered in breast
tumor
hsa-miR-4685-3p47335754discovered in breast
tumor
hsa-miR-4685-5p47345755discovered in breast
tumor
hsa-miR-468647355756discovered in breast
tumor
hsa-miR-4687-3p47365757discovered in breast
tumor
hsa-miR-4687-5p47375758discovered in breast
tumor
hsa-miR-468847385759discovered in breast
tumor
hsa-miR-468947395760discovered in breast
tumor
hsa-miR-4690-3p47405761discovered in breast
tumor
hsa-miR-4690-5p47415762discovered in breast
tumor
hsa-miR-4691-3p47425763discovered in breast
tumor
hsa-miR-4691-5p47435764discovered in breast
tumor
hsa-miR-469247445765discovered in breast
tumor
hsa-miR-4693-3p47455766discovered in breast
tumor
hsa-miR-4693-5p47465767discovered in breast
tumor
hsa-miR-4694-3p47475768discovered in breast
tumor
hsa-miR-4694-5p47485769discovered in breast
tumor
hsa-miR-4695-3p47495770discovered in breast
tumor
hsa-miR-4695-5p47505771discovered in breast
tumor
hsa-miR-469647515772discovered in breast
tumor
hsa-miR-4697-3p47525773discovered in breast
tumor
hsa-miR-4697-5p47535774discovered in breast
tumor
hsa-miR-469847545775discovered in breast
tumor
hsa-miR-4699-3p47555776discovered in breast
tumor
hsa-miR-4699-5p47565777discovered in breast
tumor
hsa-miR-4700-3p47575778discovered in breast
tumor
hsa-miR-4700-5p47585779discovered in breast
tumor
hsa-miR-4701-3p47595780discovered in breast
tumor
hsa-miR-4701-5p47605781discovered in breast
tumor
hsa-miR-4703-3p47615782discovered in breast
tumor
hsa-miR-4703-5p47625783discovered in breast
tumor
hsa-miR-4704-3p47635784discovered in breast
tumor
hsa-miR-4704-5p47645785discovered in breast
tumor
hsa-miR-470547655786discovered in breast
tumor
hsa-miR-470647665787discovered in breast
tumor
hsa-miR-4707-3p47675788discovered in breast
tumor
hsa-miR-4707-5p47685789discovered in breast
tumor
hsa-miR-4708-3p47695790discovered in breast
tumor
hsa-miR-4708-5p47705791discovered in breast
tumor
hsa-miR-4709-3p47715792discovered in breast
tumor
hsa-miR-4709-5p47725793discovered in breast
tumor
hsa-miR-471047735794discovered in breast
tumor
hsa-miR-4711-3p47745795discovered in breast
tumor
hsa-miR-4711-5p47755796discovered in breast
tumor
hsa-miR-4712-3p47765797discovered in breast
tumor
hsa-miR-4712-5p47775798discovered in breast
tumor
hsa-miR-4713-3p47785799discovered in breast
tumor
hsa-miR-4713-5p47795800discovered in breast
tumor
hsa-miR-4714-3p47805801discovered in breast
tumor
hsa-miR-4714-5p47815802discovered in breast
tumor
hsa-miR-4715-3p47825803discovered in breast
tumor
hsa-miR-4715-5p47835804discovered in breast
tumor
hsa-miR-4716-3p47845805discovered in breast
tumor
hsa-miR-4716-5p47855806discovered in breast
tumor
hsa-miR-4717-3p47865807discovered in breast
tumor
hsa-miR-4717-5p47875808discovered in breast
tumor
hsa-miR-471847885809discovered in breast
tumor
hsa-miR-471947895810discovered in breast
tumor
hsa-miR-4720-3p47905811discovered in breast
tumor
hsa-miR-4720-5p47915812discovered in breast
tumor
hsa-miR-472147925813discovered in breast
tumor
hsa-miR-4722-3p47935814discovered in breast
tumor
hsa-miR-4722-5p47945815discovered in breast
tumor
hsa-miR-4723-3p47955816discovered in breast
tumor
hsa-miR-4723-5p47965817discovered in breast
tumor
hsa-miR-4724-3p47975818discovered in breast
tumor
hsa-miR-4724-5p47985819discovered in breast
tumor
hsa-miR-4725-3p47995820discovered in breast
tumor
hsa-miR-4725-5p48005821discovered in breast
tumor
hsa-miR-4726-3p48015822discovered in breast
tumor
hsa-miR-4726-5p48025823discovered in breast
tumor
hsa-miR-4727-3p48035824discovered in breast
tumor
hsa-miR-4727-5p48045825discovered in breast
tumor
hsa-miR-4728-3p48055826discovered in breast
tumor
hsa-miR-4728-5p48065827discovered in breast
tumor
hsa-miR-472948075828discovered in breast
tumor
hsa-miR-473048085829discovered in breast
tumor
hsa-miR-4731-3p48095830discovered in breast
tumor
hsa-miR-4731-5p48105831discovered in breast
tumor
hsa-miR-4732-3p48115832discovered in breast
tumor
hsa-miR-4732-5p48125833discovered in breast
tumor
hsa-miR-4733-3p48135834discovered in breast
tumor
hsa-miR-4733-5p48145835discovered in breast
tumor
hsa-miR-473448155836discovered in breast
tumor
hsa-miR-4735-3p48165837discovered in breast
tumor
hsa-miR-4735-5p48175838discovered in breast
tumor
hsa-miR-473648185839discovered in breast
tumor
hsa-miR-473748195840discovered in breast
tumor
hsa-miR-4738-3p48205841discovered in breast
tumor
hsa-miR-4738-5p48215842discovered in breast
tumor
hsa-miR-473948225843discovered in breast
tumor
hsa-miR-4740-3p48235844discovered in breast
tumor
hsa-miR-4740-5p48245845discovered in breast
tumor
hsa-miR-474148255846discovered in breast
tumor, psoriasis
hsa-miR-4742-3p48265847discovered in breast
tumor, psoriasis
hsa-miR-4742-5p48275848discovered in breast
tumor
hsa-miR-4743-3p48285849discovered in breast
tumor
hsa-miR-4743-5p48295850discovered in breast
tumor
hsa-miR-474448305851discovered in breast
tumor
hsa-miR-4745-3p48315852discovered in breast
tumor
hsa-miR-4745-5p48325853discovered in breast
tumor
hsa-miR-4746-3p48335854discovered in breast
tumor
hsa-miR-4746-5p48345855discovered in breast
tumor
hsa-miR-4747-3p48355856discovered in breast
tumor
hsa-miR-4747-5p48365857discovered in breast
tumor
hsa-miR-474848375858discovered in breast
tumor
hsa-miR-4749-3p48385859discovered in breast
tumor
hsa-miR-4749-5p48395860discovered in breast
tumor
hsa-miR-4750-3p48405861discovered in breast
tumor
hsa-miR-4750-5p48415862discovered in breast
tumor
hsa-miR-475148425863discovered in breast
tumor
hsa-miR-475248435864discovered in breast
tumor
hsa-miR-4753-3p48445865discovered in breast
tumor
hsa-miR-4753-5p48455866discovered in breast
tumor
hsa-miR-475448465867discovered in breast
tumor
hsa-miR-4755-3p48475868discovered in breast
tumor
hsa-miR-4755-5p48485869discovered in breast
tumor
hsa-miR-4756-3p48495870discovered in breast
tumor
hsa-miR-4756-5p48505871discovered in breast
tumor
hsa-miR-4757-3p48515872discovered in breast
tumor
hsa-miR-4757-5p48525873discovered in breast
tumor
hsa-miR-4758-3p48535874discovered in breast
tumor
hsa-miR-4758-5p48545875discovered in breast
tumor
hsa-miR-475948555876discovered in breast
tumor
hsa-miR-4760-3p48565877discovered in breast
tumor
hsa-miR-4760-5p48575878discovered in breast
tumor
hsa-miR-4761-3p48585879discovered in breast
tumor
hsa-miR-4761-5p48595880discovered in breast
tumor
hsa-miR-4762-3p48605881discovered in breast
tumor
hsa-miR-4762-5p48615882discovered in breast
tumor
hsa-miR-4763-3p48625883discovered in breast
tumor
hsa-miR-4763-5p48635884discovered in breast
tumor
hsa-miR-4764-3p48645885discovered in breast
tumor
hsa-miR-4764-5p48655886discovered in breast
tumor
hsa-miR-476548665887discovered in breast
tumor
hsa-miR-4766-3p48675888discovered in breast
tumor
hsa-miR-4766-5p48685889discovered in breast
tumor
hsa-miR-476748695890discovered in breast
tumor
hsa-miR-4768-3p48705891discovered in breast
tumor
hsa-miR-4768-5p48715892discovered in breast
tumor
hsa-miR-4769-3p48725893discovered in breast
tumor
hsa-miR-4769-5p48735894discovered in breast
tumor
hsa-miR-477048745895discovered in breast
tumor
hsa-miR-477148755896discovered in breast
tumor
hsa-miR-4772-3p48765897discovered in breastenergy
tumor, bloodmetabolism/
monoclear cellsobesity
hsa-miR-4772-5p48775898discovered in breastenergy
tumor, bloodmetabolism/
monoclear cellsobesity
hsa-miR-477348785899discovered in breast
tumor
hsa-miR-4774-3p48795900discovered in breast
tumor and
Lymphoblastic
leukemia
hsa-miR-4774-5p48805901discovered in breast
tumor and
Lymphoblastic
leukemia
hsa-miR-477548815902discovered in breast
tumor
hsa-miR-4776-3p48825903discovered in breast
tumor
hsa-miR-4776-5p48835904discovered in breast
tumor
hsa-miR-4777-3p48845905discovered in breast
tumor
hsa-miR-4777-5p48855906discovered in breast
tumor
hsa-miR-4778-3p48865907discovered in breast
tumor
hsa-miR-4778-5p48875908discovered in breast
tumor
hsa-miR-477948885909discovered in breast
tumor
hsa-miR-478048895910discovered in breast
tumor
hsa-miR-4781-3p48905911discovered in breast
tumor
hsa-miR-4781-5p48915912discovered in breast
tumor
hsa-miR-4782-3p48925913discovered in breast
tumor
hsa-miR-4782-5p48935914discovered in breast
tumor
hsa-miR-4783-3p48945915discovered in breast
tumor
hsa-miR-4783-5p48955916discovered in breast
tumor
hsa-miR-478448965917discovered in breast
tumor
hsa-miR-478548975918discovered in breast
tumor
hsa-miR-4786-3p48985919discovered in breast
tumor
hsa-miR-4786-5p48995920discovered in breast
tumor
hsa-miR-4787-3p49005921discovered in breast
tumor
hsa-miR-4787-5p49015922discovered in breast
tumor
hsa-miR-478849025923discovered in breast
tumor
hsa-miR-4789-3p49035924discovered in breast
tumor
hsa-miR-4789-5p49045925discovered in breast
tumor
hsa-miR-4790-3p49055926discovered in breast
tumor
hsa-miR-4790-5p49065927discovered in breast
tumor
hsa-miR-479149075928discovered in breast
tumor
hsa-miR-479249085929discovered in breast
tumor
hsa-miR-4793-3p49095930discovered in breast
tumor
hsa-miR-4793-5p49105931discovered in breast
tumor
hsa-miR-479449115932discovered in breast
tumor
hsa-miR-4795-3p49125933discovered in breast
tumor
hsa-miR-4795-5p49135934discovered in breast
tumor
hsa-miR-4796-3p49145935discovered in breast
tumor
hsa-miR-4796-5p49155936discovered in breast
tumor
hsa-miR-4797-3p49165937discovered in breast
tumor
hsa-miR-4797-5p49175938discovered in breast
tumor
hsa-miR-4798-3p49185939discovered in breast
tumor
hsa-miR-4798-5p49195940discovered in breast
tumor
hsa-miR-4799-3p49205941discovered in breast
tumor
hsa-miR-4799-5p49215942discovered in breast
tumor
hsa-miR-4800-3p49225943discovered in breast
tumor
hsa-miR-4800-5p49235944discovered in breast
tumor
hsa-miR-480149245945discovered in breast
tumor
hsa-miR-4802-3p49255946discovered in breast
tumor, psoriasis
hsa-miR-4802-5p49265947discovered in breast
tumor, psoriasis
hsa-miR-480349275948discovered in breast
tumor
hsa-miR-4804-3p49285949discovered in breast
tumor
hsa-miR-4804-5p49295950discovered in breast
tumor
hsa-miR-483-3p49305951aderonocorticaloncogenic
carcinoma,
rectal/pancreatic
cancer,
proliferation of
wounded
epithelial cells
hsa-miR-483-5p49315952cartilageaderonocorticalangiogenesis
(chondrocyte), fetalcarcinoma
brain
hsa-miR-48449325953mitochondrial
network
hsa-miR-485-3p49335954
hsa-miR-485-5p49345955ovarian epithelial
tumor
hsa-miR-486-3p49355956erythroid cellsvarious cancers
hsa-miR-486-5p49365957stem cells (adipose)various cancers
hsa-miR-487a49375958laryngeal
carcinoma
hsa-miR-487b49385959neuroblastoma, pulmonary
carcinogenesis
hsa-miR-488-3p49395960prostate cancer,
others
hsa-miR-488-5p49405961prostate cancer,
others
hsa-miR-48949415962mesenchymal stemosteogenesis
cells
hsa-miR-490-3p49425963neuroblastoma,
terine leiomyoma
(ULM)/muscle
hsa-miR-490-5p49435964neuroblastoma,
terine leiomyoma
(ULM)/muscle
hsa-miR-491-3p49445965various cancers,pro-apoptosis
brain disease
hsa-miR-491-5p49455966various cancers,pro-apoptosis
brain disease
hsa-miR-49249465967
hsa-miR-493-3p49475968myeloid cells,
pancreas (islet)
hsa-miR-493-5p49485969myeloid cells,
pancreas (islet)
hsa-miR-49449495970epithelial cellsvarious cancerscell cycle
hsa-miR-495-3p49505971plateletvarious cancers
(gastric, MLL
leukemia,
pancreatic etc)
and inflammation
hsa-miR-495-5p49515972plateletvarious cancers
(gastric, MLL
leukemia,
pancreatic etc)
and inflammation
hsa-miR-49649525973Blood
hsa-miR-497-3p49535974various cancerstumor
(breast,supressor/pro-
colorectal, etc)apoptosis
hsa-miR-497-5p49545975various cancerstumor
(breast,supressor/pro-
colorectal, etc)apoptosis
hsa-miR-49849555976autoimmuno (e.g.
rheumatoid
arthritis)
hsa-miR-4999-3p49565977
hsa-miR-4999-5p49575978
hsa-miR-499a-3p49585979heart, cardiac stemcardiovascularcardiomyocyte
cellsdiseasedifferentiation
hsa-miR-499a-5p49595980heart, cardiac stemcardiovascularcardiomyocyte
cellsdiseasedifferentiation
hsa-miR-499b-3p49605981heart, cardiac stemcardiovascularcardiomyocyte
cellsdiseasedifferentiation
hsa-miR-499b-5p49615982heart, cardiac stemcardiovascularcardiomyocyte
cellsdiseasedifferentiation
hsa-miR-5000-3p49625983discovered in
lymphoblastic
leukaemia
hsa-miR-5000-5p49635984discovered in
lymphoblastic
leukaemia
hsa-miR-5001-3p49645985
hsa-miR-5001-5p49655986
hsa-miR-5002-3p49665987
hsa-miR-5002-5p49675988
hsa-miR-5003-3p49685989
hsa-miR-5003-5p49695990
hsa-miR-5004-3p49705991
hsa-miR-5004-5p49715992
hsa-miR-5006-3p49725993discovered in
lymphoblastic
leukaemia
hsa-miR-5006-5p49735994discovered in
lymphoblastic
leukaemia
hsa-miR-5007-3p49745995
hsa-miR-5007-5p49755996
hsa-miR-5008-3p49765997
hsa-miR-5008-5p49775998
hsa-miR-5009-3p49785999
hsa-miR-5009-5p49796000
hsa-miR-500a-3p49806001
hsa-miR-500a-5p49816002
hsa-miR-500b49826003Blood (plasma)
hsa-miR-5010-3p49836004abnormal skin
(psoriasis)
hsa-miR-5010-5p49846005abnormal skin
(psoriasis)
hsa-miR-5011-3p49856006
hsa-miR-5011-5p49866007
hsa-miR-501-3p49876008
hsa-miR-501-5p49886009
hsa-miR-502-3p49896010various cancers
(hepatocellular,
ovarian, breast)
hsa-miR-502-5p49906011various cancers
(hepatocellular,
ovarian, breast)
hsa-miR-503-3p49916012ovary
hsa-miR-503-5p49926013ovary
hsa-miR-50449936014glioblastoma
hsa-miR-504749946015
hsa-miR-505-3p49956016breast cancer
hsa-miR-505-5p49966017breast cancer
hsa-miR-506-3p49976018various cancers
hsa-miR-506-5p49986019various cancers
hsa-miR-50749996020
hsa-miR-508-3p50006021renal cell
carcinoma
hsa-miR-508-5p50016022endothelial
progenitor cells
(EPCs)
hsa-miR-508750026023
hsa-miR-508850036024
hsa-miR-5089-3p50046025
hsa-miR-5089-5p50056026
hsa-miR-509050066027
hsa-miR-509150076028
hsa-miR-509250086029
hsa-miR-509350096030
hsa-miR-509-3-5p50106031testis
hsa-miR-509-3p50116032renal cell
carcinoma, brain
disease
hsa-miR-509450126033
hsa-miR-509550136034cervical cancer
hsa-miR-509-5p50146035metabolic
syndrome, brain
disease
hsa-miR-509650156036cervical cance
hsa-miR-51050166037brain
hsa-miR-510050176038discoverd in
Salivary gland
hsa-miR-51150186039dendritic cells and
macrophages
hsa-miR-512-3p50196040embryonic stem
cells, placenta
hsa-miR-512-5p50206041embryonic stem
cells, placenta,
hsa-miR-513a-3p50216042lung carcinoma
hsa-miR-513a-5p50226043endothelial cells
hsa-miR-513b50236044follicular
lymphoma
hsa-miR-513c-3p50246045
hsa-miR-513c-5p50256046
hsa-miR-514a-3p50266047
hsa-miR-514a-5p50276048
hsa-miR-514b-3p50286049various cancer
cells
hsa-miR-514b-5p50296050various cancer
cells
hsa-miR-515-3p50306051
hsa-miR-515-5p50316052placenta
hsa-miR-516a-3p50326053frontal cortex
hsa-miR-516a-5p50336054placenta
hsa-miR-516b-3p50346055
hsa-miR-516b-5p50356056
hsa-miR-517-5p50366057placenta
hsa-miR-517a-3p50376058placenta
hsa-miR-517b-3p50386059placenta
hsa-miR-517c-3p50396060placenta
hsa-miR-518650406061discovered in
lymphoblastic
leukaemia
hsa-miR-5187-3p50416062discovered in
lymphoblastic
leukaemia, skin
(psoriasis)
hsa-miR-5187-5p50426063discovered in
lymphoblastic
leukaemia, skin
(psoriasis)
hsa-miR-518850436064discovered in
lymphoblastic
leukaemia
hsa-miR-518950446065discovered in
lymphoblastic
leukaemia
hsa-miR-518a-3p50456066HCC
hsa-miR-518a-5p50466067various cancer
cells
hsa-miR-518b50476068placentaHCCcell cycle
progression
hsa-miR-518c-3p50486069placenta
hsa-miR-518c-5p50496070placenta
hsa-miR-518d-3p50506071
hsa-miR-518d-5p50516072
hsa-miR-518e-3p50526073HCCcell cycle
progression
hsa-miR-518e-5p50536074HCCcell cycle
progression
hsa-miR-518f-3p50546075placenta
hsa-miR-518f-5p50556076placenta
hsa-miR-519050566077discovered in
lymphoblastic
leukaemia
hsa-miR-519150576078discovered in
lymphoblastic
leukaemia
hsa-miR-519250586079discovered in
lymphoblastic
leukaemia
hsa-miR-519350596080discovered in
lymphoblastic
leukaemia
hsa-miR-519450606081discovered in
lymphoblastic
leukaemia
hsa-miR-5195-3p50616082discovered in
lymphoblastic
leukaemia
hsa-miR-5195-5p50626083discovered in
lymphoblastic
leukaemia
hsa-miR-5196-3p50636084discovered in
lymphoblastic
leukaemia
hsa-miR-5196-5p50646085discovered in
lymphoblastic
leukaemia
hsa-miR-5197-3p50656086discovered in
lymphoblastic
leukaemia
hsa-miR-5197-5p50666087discovered in
lymphoblastic
leukaemia
hsa-miR-519a-3p50676088placentaHCC
hsa-miR-519a-5p50686089placentaHCC
hsa-miR-519b-3p50696090breast cancer
hsa-miR-519b-5p50706091breast cancer
hsa-miR-519c-3p50716092
hsa-miR-519c-5p50726093
hsa-miR-519d50736094placenta
hsa-miR-519e-3p50746095placenta
hsa-miR-519e-5p50756096placenta
hsa-miR-520a-3p50766097placenta
hsa-miR-520a-5p50776098placenta
hsa-miR-520b50786099breast cancer
hsa-miR-520c-3p50796100gastric cancer,
breast tumor
hsa-miR-520c-5p50806101breast tumor
hsa-miR-520d-3p50816102various cancer
cells
hsa-miR-520d-5p50826103various cancer
cells
hsa-miR-520e50836104hepatomatomor
suppressor
hsa-miR-520f50846105breast cancer
hsa-miR-520g50856106HCC, bladder
cancer, breast
cancer
hsa-miR-520h50866107placental specific
hsa-miR-52150876108prostate cancer
hsa-miR-522-3p50886109HCC
hsa-miR-522-5p50896110HCC
hsa-miR-523-3p50906111
hsa-miR-523-5p50916112
hsa-miR-524-3p50926113colon cancer stem
cells
hsa-miR-524-5p50936114placental specificgliomas
hsa-miR-525-3p50946115placental specificHCC
hsa-miR-525-5p50956116placental specific
hsa-miR-526a50966117placental specific
hsa-miR-526b-3p50976118placental specific
hsa-miR-526b-5p50986119placental specific
hsa-miR-52750996120
hsa-miR-532-3p51006121ALL
hsa-miR-532-5p51016122ALL
hsa-miR-539-3p51026123
hsa-miR-539-5p51036124
hsa-miR-541-3p51046125
hsa-miR-541-5p51056126
hsa-miR-542-3p51066127monocytes
hsa-miR-542-5p51076128basal cell
carcinoma,
neuroblastoma
hsa-miR-54351086129
hsa-miR-544a51096130osteocarcoma
hsa-miR-544b51106131osteocarcoma
hsa-miR-545-3p51116132
hsa-miR-545-5p51126133rectal cancer
hsa-miR-54851136134
hsa-miR-548-3p51146135
hsa-miR-548-5p51156136
hsa-miR-548a51166137identified in
colorectal
microRNAome
hsa-miR-548a-3p51176138identified in
colorectal
microRNAome
hsa-miR-548a-5p51186139identified in
colorectal
microRNAome
hsa-miR-548aa51196140identified in
cervical tumor
hsa-miR-548ab51206141discovered in B-
cells
hsa-miR-548ac51216142discovered in B-
cells
hsa-miR-548ad51226143discovered in B-
cells
hsa-miR-548ae51236144discovered in B-
cells
hsa-miR-548ag51246145discovered in B-
cells
hsa-miR-548ah-3p51256146discovered in B-
cells
hsa-miR-548ah-5p51266147discovered in B-
cells
hsa-miR-548ai51276148discovered in B-
cells
hsa-miR-548aj-3p51286149discovered in B-
cells
hsa-miR-548aj-5p51296150discovered in B-
cells
hsa-miR-548ak51306151discovered in B-
cells
hsa-miR-548al51316152discovered in B-
cells
hsa-miR-548am-3p51326153discovered in B-
cells
hsa-miR-548am-5p51336154discovered in B-
cells
hsa-miR-548an51346155discovered in B-
cells
hsa-miR-548ao-3p51356156
hsa-miR-548ao-5p51366157
hsa-miR-548ap-3p51376158
hsa-miR-548ap-5p51386159
hsa-miR-548aq-3p51396160
hsa-miR-548aq-5p51406161
hsa-miR-548ar-3p51416162
hsa-miR-548ar-5p51426163
hsa-miR-548as-3p51436164
hsa-miR-548as-5p51446165
hsa-miR-548at-3p51456166prostate cancer
hsa-miR-548at-5p51466167prostate cancer
hsa-miR-548au-3p51476168
hsa-miR-548au-5p51486169
hsa-miR-548av-3p51496170
hsa-miR-548av-5p51506171
hsa-miR-548aw51516172prostate cancer
hsa-miR-548ay-3p51526173discovered in
abnormal skin
(psoriasis)
hsa-miR-548ay-5p51536174discovered in
abnormal skin
(psoriasis)
hsa-miR-548az-3p51546175discovered in
abnormal skin
(psoriasis)
hsa-miR-548az-5p51556176discovered in
abnormal skin
(psoriasis)
hsa-miR-548b-3p51566177identified in
colorectal
microRNAome
hsa-miR-548b-5p51576178immune cells,
frontal cortex
hsa-miR-548c-3p51586179identified in
colorectal
microRNAome
hsa-miR-548c-5p51596180immune cells,
frontal cortex
hsa-miR-548d-3p51606181identified in
colorectal
microRNAome
hsa-miR-548d-5p51616182identified in
colorectal
microRNAome
hsa-miR-548e51626183embryonic stem
cells
hsa-miR-548f51636184embryonic stem
cells
hsa-miR-548g-3p51646185embryonic stem
cells
hsa-miR-548g-5p51656186embryonic stem
cells
hsa-miR-548h-3p51666187embryonic stem
cells
hsa-miR-548h-5p51676188embryonic stem
cells
hsa-miR-548i51686189embryonic stem
cells, immune cells
hsa-miR-548j51696190immune cells
hsa-miR-548k51706191embryonic stem
cells
hsa-miR-548151716192embryonic stem
cells
hsa-miR-548m51726193embryonic stem
cells
hsa-miR-548n51736194embryonic stem
cells, immune cells
hsa-miR-548o-3p51746195embryonic stem
cells
hsa-miR-548o-5p51756196embryonic stem
cells
hsa-miR-548p51766197embryonic stem
cells
hsa-miR-548q51776198ovarian cancer
cells
hsa-miR-548s51786199discovered in the
melanoma
MicroRNAome
hsa-miR-548t-3p51796200discovered in the
melanoma
MicroRNAome
hsa-miR-548t-5p51806201discovered in the
melanoma
MicroRNAome
hsa-miR-548u51816202discovered in the
melanoma
MicroRNAome
hsa-miR-548w51826203discovered in the
melanoma
MicroRNAome
hsa-miR-548y51836204
hsa-miR-548z51846205discovered in
cervical tumor
hsa-miR-549a51856206discovered in a
colorectal
MicroRNAome
hsa-miR-550a-3-5p51866207Hepatocellular
Carcinoma
hsa-miR-550a-3p51876208Hepatocellular
Carcinoma
hsa-miR-550a-5p51886209Hepatocellular
Carcinoma
hsa-miR-550b-2-5p51896210discovered in
cervical tumor
hsa-miR-550b-3p51906211discovered in
cervical tumor
hsa-miR-551a51916212gastric cancer
hsa-miR-551b-3p51926213hepatocytes
hsa-miR-551b-5p51936214hepatocytes
hsa-miR-55251946215discovered in a
colorectal
MicroRNAome
hsa-miR-55351956216discovered in a
colorectal
MicroRNAome
hsa-miR-55451966217discovered in a
colorectal
MicroRNAome
hsa-miR-55551976218discovered in a
colorectal
MicroRNAome
hsa-miR-556-3p51986219discovered in a
colorectal
MicroRNAome
hsa-miR-556-5p51996220discovered in a
colorectal
MicroRNAome
hsa-miR-55752006221liver (hepatocytes)
hsa-miR-5571-3p52016222discoveredd in
Salivary gland
hsa-miR-5571-5p52026223discoveredd in
Salivary gland
hsa-miR-557252036224discoveredd in
Salivary gland
hsa-miR-5579-3p52046225
hsa-miR-5579-5p52056226
hsa-miR-55852066227neuroblastoma
hsa-miR-5580-3p52076228
hsa-miR-5580-5p52086229
hsa-miR-5581-3p52096230
hsa-miR-5581-5p52106231
hsa-miR-5582-3p52116232
hsa-miR-5582-5p52126233
hsa-miR-5583-3p52136234
hsa-miR-5583-5p52146235
hsa-miR-5584-3p52156236
hsa-miR-5584-5p52166237
hsa-miR-5585-3p52176238
hsa-miR-5585-5p52186239
hsa-miR-5586-3p52196240
hsa-miR-5586-5p52206241
hsa-miR-5587-3p52216242
hsa-miR-5587-5p52226243
hsa-miR-5588-3p52236244
hsa-miR-5588-5p52246245
hsa-miR-5589-3p52256246
hsa-miR-5589-5p52266247
hsa-miR-55952276248
hsa-miR-5590-3p52286249
hsa-miR-5590-5p52296250
hsa-miR-5591-3p52306251
hsa-miR-5591-5p52316252
hsa-miR-561-3p52326253multiple myeloma
hsa-miR-561-5p52336254multiple myeloma
hsa-miR-56252346255
hsa-miR-56352356256discovered in a
colorectal
MicroRNAome
hsa-miR-56452366257Chronic myeloid
leukemia
hsa-miR-56652376258MALT
lymphoma/lymphocyte
hsa-miR-56752386259colorectal cancer
hsa-miR-56852396260discovered in a
colorectal
MicroRNAome
hsa-miR-568052406261Associated with
metastatic
prostate cancer
hsa-miR-5681a52416262Associated with
metastatic
prostate cancer
hsa-miR-5681b52426263Associated with
metastatic
prostate cancer
hsa-miR-568252436264Associated with
metastatic
prostate cancer
hsa-miR-568352446265Associated with
metastatic
prostate cancer
hsa-miR-568452456266Associated with
metastatic
prostate cancer
hsa-miR-568552466267Associated with
metastatic
prostate cancer
hsa-miR-568652476268Associated with
metastatic
prostate cancer
hsa-miR-568752486269Associated with
metastatic
prostate cancer
hsa-miR-568852496270Associated with
metastatic
prostate cancer
hsa-miR-568952506271Associated with
metastatic
prostate cancer
hsa-miR-56952516272
hsa-miR-569052526273Associated with
metastatic
prostate cancer
hsa-miR-569152536274Associated with
metastatic
prostate cancer
hsa-miR-5692a52546275Associated with
metastatic
prostate cancer
hsa-miR-5692b52556276Associated with
metastatic
prostate cancer
hsa-miR-5692c52566277Associated with
metastatic
prostate cancer
hsa-miR-569352576278Associated with
metastatic
prostate cancer
hsa-miR-569452586279Associated with
metastatic
prostate cancer
hsa-miR-569552596280Associated with
metastatic
prostate cancer
hsa-miR-569652606281Associated with
metastatic
prostate cancer
hsa-miR-569752616282Associated with
metastatic
prostate cancer
hsa-miR-569852626283Associated with
metastatic
prostate cancer
hsa-miR-569952636284Associated with
metastatic
prostate cancer
hsa-miR-570052646285Associated with
metastatic
prostate cancer
hsa-miR-570152656286Associated with
metastatic
prostate cancer
hsa-miR-570252666287Associated with
metastatic
prostate cancer
hsa-miR-570352676288Associated with
metastatic
prostate cancer
hsa-miR-570-3p52686289follicular
lymphoma
hsa-miR-570452696290Associated with
metastatic
prostate cancer
hsa-miR-570552706291Associated with
metastatic
prostate cancer
hsa-miR-570-5p52716292follicular
lymphoma
hsa-miR-570652726293Associated with
metastatic
prostate cancer
hsa-miR-570752736294Associated with
metastatic
prostate cancer
hsa-miR-570852746295Associated with
metastatic
prostate cancer
hsa-miR-57152756296frontal cortex
hsa-miR-57252766297circulatingbasal cell
microRNA (incarcinoma
plasma)
hsa-miR-57352776298discovered in the
colorectal
MicroRNAome
hsa-miR-573952786299endothelial cells
hsa-miR-574-3p52796300blood (myeloidfollicular
cells)lymphoma
hsa-miR-574-5p52806301semen
hsa-miR-57552816302gastric cancer
hsa-miR-576-3p52826303discovered in a
colorectal
MicroRNAome
hsa-miR-576-5p52836304cartilage/
chondrocyte
hsa-miR-57752846305discovered in a
colorectal
MicroRNAome
hsa-miR-57852856306discovered in a
colorectal
MicroRNAome
hsa-miR-578752866307fibroblast
hsa-miR-57952876308
hsa-miR-58052886309breast cancer
hsa-miR-58152896310liver (hepatocytes)
hsa-miR-582-3p52906311cartilage/chondrocytebladder cancer
hsa-miR-582-5p52916312bladder cancer
hsa-miR-58352926313rectal cancer cells
hsa-miR-584-3p52936314tumor cells
(follicular
lymphoma, rectal
cancer cells)
hsa-miR-584-5p52946315tumor cells
(follicular
lymphoma, rectal
cancer cells)
hsa-miR-58552956316oral squamous
cell carcinoma
hsa-miR-58652966317discovered in a
colorectal
MicroRNAome
hsa-miR-58752976318discovered in a
colorectal
MicroRNAome
hsa-miR-58852986319discovered in a
colorectal
MicroRNAome
hsa-miR-589-3p52996320mesothelial cells
hsa-miR-589-5p53006321mesothelial cells
hsa-miR-590-3p53016322cardiomyocytesCell cycle
progression
hsa-miR-590-5p53026323cardiomyocytesCell cycle
progression
hsa-miR-59153036324neuroblastoma
hsa-miR-59253046325hepatocellular
carcinoma
hsa-miR-593-3p53056326esophageal cancer
hsa-miR-593-5p53066327esophageal cancer
hsa-miR-59553076328heart failure
hsa-miR-59653086329ependymoma,
cancers
hsa-miR-59753096330discovered in a
colorectal
MicroRNAome
hsa-miR-59853106331Blood
(lymphocytes)
hsa-miR-59953116332Multiple sclerosis
hsa-miR-60053126333discovered in a
colorectal
MicroRNAome
hsa-miR-60153136334various cancers
(colonrectal,
gastric)
hsa-miR-60253146335oocyte
hsa-miR-60353156336
hsa-miR-60453166337discovered in a
colorectal
MicroRNAome
hsa-miR-60553176338discovered in a
colorectal
MicroRNAome
hsa-miR-60653186339discovered in a
colorectal
MicroRNAome
hsa-miR-606853196340discovered in
endothelial cells
hsa-miR-606953206341discovered in
endothelial cells
hsa-miR-60753216342discovered in a
colorectal
MicroRNAome
hsa-miR-607053226343discovered in a
colorectal
MicroRNAome
hsa-miR-607153236344discovered in
endothelial cells
hsa-miR-607253246345discovered in
endothelial cells
hsa-miR-607353256346discovered in
endothelial cells
hsa-miR-607453266347discovered in
endothelial cells
hsa-miR-607553276348discovered in
endothelial cells
hsa-miR-607653286349discovered in
endothelial cells
hsa-miR-607753296350discovered in
endothelial cells
hsa-miR-607853306351discovered in
endothelial cells
hsa-miR-607953316352discovered in
endothelial cells
hsa-miR-60853326353various cancers
hsa-miR-608053336354discovered in
endothelial cells
hsa-miR-608153346355discovered in
endothelial cells
hsa-miR-608253356356discovered in
endothelial cells
hsa-miR-608353366357discovered in
endothelial cells
hsa-miR-608453376358discovered in
endothelial cells
hsa-miR-608553386359discovered in
endothelial cells
hsa-miR-608653396360embryonic stem
cells
hsa-miR-608753406361embryonic stem
cells
hsa-miR-608853416362embryonic stem
cells
hsa-miR-608953426363embryonic stem
cells
hsa-miR-60953436364discovered in a
colorectal
MicroRNAome
hsa-miR-609053446365embryonic stem
cells
hsa-miR-61053456366gastric cancer
hsa-miR-61153466367Renal cell
carcinoma
hsa-miR-61253476368AM leukemia
hsa-miR-612453486369
hsa-miR-612553496370
hsa-miR-612653506371
hsa-miR-612753516372
hsa-miR-612853526373
hsa-miR-612953536374
hsa-miR-61353546375lipid metabollism
hsa-miR-613053556376
hsa-miR-613153566377
hsa-miR-613253576378
hsa-miR-613353586379
hsa-miR-613453596380
hsa-miR-61453606381circulating
micrRNAs (in
Plasma)
hsa-miR-615-3p53616382
hsa-miR-615-5p53626383
hsa-miR-616-3p53636384prostate cancer
hsa-miR-616553646385Pro-apoptotic
factor
hsa-miR-616-5p53656386prostate cancer
hsa-miR-61753666387
hsa-miR-61853676388
hsa-miR-61953686389discovered in a
colorectal
MicroRNAome
hsa-miR-62053696390discovered in a
colorectal
MicroRNAome
hsa-miR-62153706391
hsa-miR-62253716392
hsa-miR-62353726393
hsa-miR-624-3p53736394chondrocyte
hsa-miR-624-5p53746395chondrocyte
hsa-miR-625-3p53756396liver (hepatocytes), circulatingvarious cancers
(blood)
hsa-miR-625-5p53766397liver (hepatocytes), circulatingvarious cancers
(blood)
hsa-miR-62653776398discovered in the
colorectal
MicroRNAome
hsa-miR-62753786399colorectal cancer
hsa-miR-628-3p53796400neuroblastoma
hsa-miR-628-5p53806401neuroblastoma
hsa-miR-629-3p53816402B-lineage ALL, T
cell lupus,
RCC/kidney
hsa-miR-629-5p53826403B-lineage ALL, T
cell lupus,
RCC/kidney
hsa-miR-63053836404chondrocytesrectal cancer
hsa-miR-63153846405discovered in the
colorectal
MicroRNAom
hsa-miR-63253856406myelodysplastic
syndromes
hsa-miR-63353866407multiple sclerosis
hsa-miR-63453876408cartilage/
chondrocyte
hsa-miR-63553886409discovered in the
colorectal
MicroRNAome
hsa-miR-63653896410myelodysplastic
syndromes
hsa-miR-63753906411discovered in the
colorectal
MicroRNAome
hsa-miR-63853916412Lupus nephritis,
basal cell
carcinoma
hsa-miR-63953926413discovered in the
colorectal
MicroRNAome
hsa-miR-64053936414Chronic
lymphocytic
leukemia
hsa-miR-64153946415cartilage/
chondrocyte
hsa-miR-642a-3p53956416adipocyte
hsa-miR-642a-5p53966417discovered in the
colorectal
MicroRNAome
hsa-miR-642b-3p53976418discovered in a
cervial tumo
hsa-miR-642b-5p53986419discovered in a
cervial tumo
hsa-miR-64353996420discovered in the
colorectal
MicroRNAome
hsa-miR-644a54006421
hsa-miR-64554016422ovarian cancer
hsa-miR-64654026423
hsa-miR-64754036424prostate and lung
cancer
hsa-miR-64854046425circulating
micrRNAs (in
Plasma)
hsa-miR-64954056426Serum
hsa-miR-6499-3p54066427discovered in
abnormal skin
(psoriasis)
hsa-miR-6499-5p54076428discovered in
abnormal skin
(psoriasis)
hsa-miR-65054086429melanoma
hsa-miR-6500-3p54096430discovered in
abnormal skin
(psoriasis)
hsa-miR-6500-5p54106431discovered in
abnormal skin
(psoriasis)
hsa-miR-6501-3p54116432discovered in
abnormal skin
(psoriasis)
hsa-miR-6501-5p54126433discovered in
abnormal skin
(psoriasis)
hsa-miR-6502-3p54136434discovered in
abnormal skin
(psoriasis)
hsa-miR-6502-5p54146435discovered in
abnormal skin
(psoriasis)
hsa-miR-6503-3p54156436discovered in
abnormal skin
(psoriasis)
hsa-miR-6503-5p54166437discovered in
abnormal skin
(psoriasis)
hsa-miR-6504-3p54176438discovered in
abnormal skin
(psoriasis)
hsa-miR-6504-5p54186439discovered in
abnormal skin
(psoriasis)
hsa-miR-6505-3p54196440discovered in
abnormal skin
(psoriasis)
hsa-miR-6505-5p54206441discovered in
abnormal skin
(psoriasis)
hsa-miR-6506-3p54216442discovered in
abnormal skin
(psoriasis)
hsa-miR-6506-5p54226443discovered in
abnormal skin
(psoriasis)
hsa-miR-6507-3p54236444discovered in
abnormal skin
(psoriasis)
hsa-miR-6507-5p54246445discovered in
abnormal skin
(psoriasis)
hsa-miR-6508-3p54256446discovered in
abnormal skin
(psoriasis)
hsa-miR-6508-5p54266447discovered in
abnormal skin
(psoriasis)
hsa-miR-6509-3p54276448discovered in
abnormal skin
(psoriasis)
hsa-miR-6509-5p54286449discovered in
abnormal skin
(psoriasis)
hsa-miR-65154296450discovered in thelung cancer
colorectal
MicroRNAome
hsa-miR-6510-3p54306451discovered in
abnormal skin
(psoriasis)
hsa-miR-6510-5p54316452discovered in
abnormal skin
(psoriasis)
hsa-miR-6511a-3p54326453discovered in
abnormal skin
(psoriasis) and
epididymis
hsa-miR-6511a-5p54336454discovered in
abnormal skin
(psoriasis) and
epididymis
hsa-miR-6511b-3p54346455discovered in
epididymis
hsa-miR-6511b-5p54356456discovered in
epididymis
hsa-miR-6512-3p54366457discovered in
abnormal skin
(psoriasis)
hsa-miR-6512-5p54376458discovered in
abnormal skin
(psoriasis)
hsa-miR-6513-3p54386459discovered in
abnormal skin
(psoriasis)
hsa-miR-6513-5p54396460discovered in
abnormal skin
(psoriasis)
hsa-miR-6514-3p54406461discovered in
abnormal skin
(psoriasis)
hsa-miR-6514-5p54416462discovered in
abnormal skin
(psoriasis)
hsa-miR-6515-3p54426463discovered in
abnormal skin
(psoriasis) and
epididymis
hsa-miR-6515-5p54436464discovered in
abnormal skin
(psoriasis) and
epididymis
hsa-miR-652-3p54446465rectal cancer cells
hsa-miR-652-5p54456466rectal cancer cells
hsa-miR-65354466467Discovered in the
colorectal
MicroRNAome
hsa-miR-654-3p54476468Discovered in the
colorectal
MicroRNAome
hsa-miR-654-5p54486469bone marrowprostate cancer
hsa-miR-65554496470
hsa-miR-65654506471various cancers
hsa-miR-65754516472oligodendrocytesdiabetes
hsa-miR-65854526473gastric cancer
hsa-miR-659-3p54536474myoblast
hsa-miR-659-5p54546475myoblast
hsa-miR-660-3p54556476myoblast
hsa-miR-660-5p54566477myoblast
hsa-miR-66154576478breast cancer
hsa-miR-66254586479endothelial
progenitor cells,
oocytes
hsa-miR-663a54596480follicular
lymphoma, Lupus
nephritis
hsa-miR-663b54606481follicular
lymphoma, Lupus
nephritis
hsa-miR-664a-3p54616482embryonic stemcomponent of
cellsSnoRNAs
hsa-miR-664a-5p54626483embryonic stemcomponent of
cellsSnoRNAs
hsa-miR-664b-3p54636484embryonic stemcomponent of
cellsSnoRNAs
hsa-miR-664b-5p54646485embryonic stemcomponent of
cellsSnoRNAs
hsa-miR-66554656486breast cancer
hsa-miR-66854666487keratinocytessenescence
hsa-miR-67054676488
hsa-miR-671-3p54686489
hsa-miR-6715a-3p54696490discovered in
epididymis
hsa-miR-6715b-3p54706491discovered in
epididymis
hsa-miR-6715b-5p54716492discovered in
epididymis
hsa-miR-671-5p54726493rectal cancer,
prolactinomas
hsa-miR-6716-3p54736494discovered in
epididymis
hsa-miR-6716-5p54746495discovered in
epididymis
hsa-miR-6717-5p54756496discovered in
epididymis
hsa-miR-6718-5p54766497discovered in
epididymis
hsa-miR-6719-3p54776498discovered in
epididymis
hsa-miR-6720-3p54786499discovered in
epididymis
hsa-miR-6721-5p54796500discovered in
epididymis
hsa-miR-6722-3p54806501discovered in
epididymis
hsa-miR-6722-5p54816502discovered in
epididymis
hsa-miR-6723-5p54826503discovered in
epididymis
hsa-miR-6724-5p54836504discovered in
epididymis
hsa-miR-675-3p54846505adrenocortical
tumor
hsa-miR-675-5p54856506adrenocortical
tumor
hsa-miR-676-3p54866507discovered in
female
reproductive tract
hsa-miR-676-5p54876508discovered in
female
reproductive tract
hsa-miR-708-3p54886509Various cancers
(lung, bladder,
pancreatic, ALL)
hsa-miR-708-5p54896510Various cancers
(lung, bladder,
pancreatic, ALL)
hsa-miR-71154906511cutaneous T-cell
lymphomas
hsa-miR-7-1-3p54916512Glioblast, brain,
prancreas
hsa-miR-71854926513blood
hsa-miR-7-2-3p54936514brain, pancreas
hsa-miR-744-3p54946515heart
hsa-miR-744-5p54956516embryonic stem
cells, heart
hsa-miR-758-3p54966517cholesterol
regulation and brain
hsa-miR-758-5p54976518cholesterol
regulation and brain
hsa-miR-75954986519
hsa-miR-7-5p54996520brain
hsa-miR-76055006521colonrectal and
breast cancer
hsa-miR-76155016522
hsa-miR-76255026523corneal epithelial
cells
hsa-miR-76455036524osteoblast
hsa-miR-76555046525rectal cancer
hsa-miR-766-3p55056526embryonic stem
cells
hsa-miR-766-5p55066527embryonic stem
cells
hsa-miR-767-3p55076528/
hsa-miR-767-5p55086529/
hsa-miR-769-3p55096530
hsa-miR-769-5p55106531
hsa-miR-770-5p55116532
hsa-miR-80255126533brain, epithelialdown symdrome
cells, hepatocytes
hsa-miR-873-3p55136534
hsa-miR-873-5p55146535
hsa-miR-87455156536cervical cancer,
lung cancer,
carcinoma
hsa-miR-875-3p55166537
hsa-miR-875-5p55176538
hsa-miR-876-3p55186539
hsa-miR-876-5p55196540
hsa-miR-877-3p55206541
hsa-miR-877-5p55216542
hsa-miR-885-3p55226543embryonic stem
cells
hsa-miR-885-5p55236544embryonic stem
cells
hsa-miR-88755246545
hsa-miR-888-3p55256546
hsa-miR-888-5p55266547
hsa-miR-88955276548
hsa-miR-89055286549epididymis
hsa-miR-891a55296550epididymisosteosarcoma
hsa-miR-891b55306551epididymis
hsa-miR-892a55316552epididymis
hsa-miR-892b55326553epididymis
hsa-miR-892c-3p55336554discovered in
epididymis
hsa-miR-892c-5p55346555discovered in
epididymis
hsa-miR-92055356556human testis
hsa-miR-92155366557human testismuscle invasive
bladder cancer
hsa-miR-92255376558human testis,multiple sclerosis,
neuronal tissuesAlcoholic liver
disease
hsa-miR-92455386559human testis
hsa-miR-92a-1-5p55396560endothelial cells
hsa-miR-92a-2-5p55406561endothelial cells
hsa-miR-92a-3p55416562endothelial cells,
CNS
hsa-miR-92b-3p55426563endothelial cells,
heart
hsa-miR-92b-5p55436564endothelial cells,
heart
hsa-miR-93355446565discovered in
cervical cancer
hsa-miR-93-3p55456566embryonic stembasal cell
cellscarcinoma
hsa-miR-93455466567discovered in
cervical cancer
hsa-miR-93555476568blood monoclearenergy
cellsmetabolism/
obesity,
medullablastoma/
neural stem cells
hsa-miR-93-5p55486569embryonic stem
cells
hsa-miR-93655496570skin
hsa-miR-937-3p55506571cervical cancer
hsa-miR-937-5p55516572cervical cancer
hsa-miR-93855526573Various cancer
cells
hsa-miR-939-3p55536574hepatocytes
hsa-miR-939-5p55546575hepatocytes
hsa-miR-9-3p55556576brainCancers and brain
diseases
hsa-miR-94055566577identified in
Cervical cancer
hsa-miR-94155576578Embryonic stem
cells
hsa-miR-94255586579lung cancer
hsa-miR-94355596580identified in
Cervical cancer
hsa-miR-94455606581various cancers
(cervical,
pancreatic,
colonrectal)
hsa-miR-9555616582various cancers
(pancreatic,
glioblastoma,
colorectal etc)
hsa-miR-9-5p55626583brainCancers and brain
disease
hsa-miR-96-3p55636584stem cellsvarious cancers
(prostate,
lymphoma, HCC,
etc) and
inflammation
hsa-miR-96-5p55646585stem cellsvarious cancers
(prostate,
lymphoma, HCC,
etc) and
inflammation
hsa-miR-98-3p55656586various cancerapoptosis
cells
hsa-miR-98-5p55666587various cancerapoptosis
cells
hsa-miR-99a-3p55676588hemapoietic cells
hsa-miR-99a-5p55686589hemapoietic cells
hsa-miR-99b-3p55696590hemapoietic cells,
embryonic stem
cells
hsa-miR-99b-5p55706591hemapoietic cells,
embryonic stem
cells

MicroRNAs that are enriched in specific types of immune cells are listed in Table 11. Furthermore, novel microRNAs are discovered in the immune cells in the art through micro-array hybridization and microtome analysis (Jima D D et al, Blood, 2010, 116:e118-e127; Vaz C et al., BMC Genomics, 2010, 11,288, the content of each of which is incorporated herein by reference in its entirety). In Table 11, “HCC” represents hepatocellular carcinoma, “ALL” stands for acute lymphoblastsic leukemia and “CLL” stands for chrominc lymphocytic leukemia.

TABLE 11
microRNAs in immune cells
mirBStissues/cells
SEQSEQwithbiological
microRNAIDIDMicroRNAsassociated diseasesfunctions/targets
hsa-let-7a-2-3p25083529embryonic steminflammatory,tumor
cells, lung,various cancerssuppressor,
myeloid cells(lung, cervical,target to c-myc
breast, pancreatic,
etc)
hsa-let-7a-3p25093530embryonic steminflammatory,tumor
cell, lung,various cancerssuppressor,
myeloid cells(lung, cervical,target to c-myc
breast, pancreatic,
etc)
hsa-let-7a-5p25103531embryonic steminflammatory,tumor
cells, lung,various cancerssuppressor,
myeloid cells(lung, cervical,target to c-myc
breast, pancreatic,
etc)
hsa-let-7c25133534dendritic cellsvarious cacnerstumor
(cervical, pancreatic,suppressor
lung, esopphageal,apoptosis
etc)(target to BCL-
x1)
hsa-let-7e-3p25163537immune cellsvarious cancer cells,tumor
autoimmunitysuppressor
TLR signal pathway
in endotoxin
tolerance
hsa-let-7e-5p25173538immune cellsassociated withtumor
various cancer cellssuppressor
hsa-let-7f-1-3p25183539immune cells (Tassociated withtumor
cells)various cancer cellssuppressor
hsa-let-7f-2-3p25193540immune cells (Tassociated withtumor
cells)various cancer cellssuppressor
hsa-let-7f-5p25203541immune cells (Tassociated withtumor
cells)various cancer cellssuppressor
hsa-let-7g-3p25213542hematopoieticvarious cancer cellstumor
cells, adipose,(lung, breast, etc)suppressor
smooth muscle(target to
cellsNFkB, LOX1)
hsa-let-7g-5p25223543hematopoieticvarious cancer cellstumor
cells, adipose,(lung, breast, etc)suppressor
smooth muscle(target to
cellsNFkB, LOX1)
hsa-let-7i-3p25233544immune cellschronic lymphocytetumor
leukimiasuppressor
hsa-let-7i-5p25243545immune cellschronic lymphocytetumor
leukimiasuppressor
hsa-miR-10a-3p25303551hematopoeiticacute myeoidoncogene, cell
cellsleukemiagrowth
hsa-miR-10a-5p25413562hematopoieticacute myeloidoncogene, cell
cellsleukemiagrowth
hsa-miR-118425513572Hematopoieticdownregulated inpredited in the
cellsoral leukoplakiaintron 22 of F8
(OLK)gene
hsa-miR-125b-1-26163637hematopoieticvarious canceroncogene, cell
3pcells(ALL, prostate,differentiation
(monocytes),HCC, etc); TLR
brain (neuron)signal pathway in
endotoxin tolerance
hsa-miR-125b-2-26173638hematopoieticvarious canceroncogene cell
3pcells(ALL, prostate,differentiation
(monocytes),HCC etc); TLR
brain (neuron)signal pathway in
endotoxin tolerance
hsa-miR-125b-26183639hematopoieticvarious canceroncogene cell
5pcells, brain(Cutaneous T celldifferentiation
(neuron)lymphomas,
prostate, HCC, etc);
TLR signal pathway
in endotoxin
tolerance
hsa-miR-127926523673monocytes
hsa-miR-130a-3p26903711lung, monocytes,various cancerspro-angiogenic
vascular(basal cell
endothelial cellscarcinoma,
HCC, ovarian, etc),
drug resistance
hsa-miR-130a-5p26913712lung, monocytes,various cancerspro-angiogenic
vasscular(basal cell
endothelial cellscarcinoma,
HCC, ovarian, etc),
drug resistance
hsa-miR-132-3p26973718brain(neuron),
immune cells
hsa-miR-132-5p26993720brain(neuron),
immune cells
hsa-miR-142-3p27203741meyloid cells,tumor
hematopoiesis,suppressor,
APC cellsimmune
response
hsa-miR-142-5p27213742meyloid cells,immune
hematopoiesis,response
APC cells
hsa-miR-143-5p27233744vascular smoothincreased in serum
muscle, T-cellsafter virus infection
hsa-miR-146a-3p27303751immune cells,associated with
hematopoiesis,CLL, TLR signal
cartilage,pathway in
endotoxin tolerance
hsa-miR-146a-5p27313752immune cells,associated with
hematopoiesis,CLL, TLR signal
cartilage,pathway in
endotoxin tolerance
hsa-miR-146b-27323753immune cellscancers (thyroidimmune
3pcarcimona)response
hsa-miR-146b-27333754embryoid bodythyroid cancer,tumor invation,
5pcellsassociated with CLLmigration
hsa-miR-147a27363757Macrophageinflammatory
response
hsa-miR-147b27373758Macrophageinflammatory
response
hsa-miR-148a-3p27383759hematopoieticassociated with
cellsCLL, T-lineage
ALL
hsa-miR-148a-5p27393760hematopoieticassociated with
cellsCLL, T-lineage
ALL
hsa-miR-150-3p27443765hematopoiticcirculating plasma
cells (lymphoid)(acute myeloid
leukemia)
hsa-miR-150-5p27453766hematopoiticcirculating plasma
cells (lymphoid)(acute myeloid
leukemia)
hsa-miR-151b27483769immune cells (B-
cells)
hsa-miR-155-3p27563777T/B cells,associated with
monocytes, breastCLL, TLR signal
pathway in
endotoxin tolerance;
upregulated in B
cell lymphoma
(CLL) and other
cancers (breast,
lung, ovarian,
cervical, colorectal,
prostate)
hsa-miR-155-5p27573778T/B cells,associated with CLL,
monocytes, breastTLR signal
pathway in
endotoxin tolerance,
upregulated in B
cell lymphoma
(CLL) and other
cancers (breast,
lung, ovarian,
cervical, colorectal,
prostate)
hsa-miR-15a-3p27593780blood,chronic lymphocytic
lymphocyte,leukemia
hematopoietic
tissues (spleen)
hsa-miR-15a-5p27603781blood,chronic lymphocytic
lymphocyte,leukemia
hematopoietic
tissues (spleen)
hsa-miR-15b-3p27613782blood,cell cycle,
lymphocyte,proliferation
hematopoietic
tissues (spleen)
hsa-miR-15b-5p27623783blood,cell cycle,
lymphocyte,proliferation
hematopoietic
tissues (spleen)
hsa-miR-16-1-3p27633784embryonic stemchronic lymphocytic
cells, blood,leukemia
hematopoietic
tissues (spleen)
hsa-miR-16-2-3p27643785blood,
lymphocyte,
hematopoietic
tissues (spleen)
hsa-miR-16-5p27653786blood,
lymphocyte,
hematopoietic
tissues
hsa-miR-181a-3p27693790glioblast,
myeloid cells,
Embryonic stem
cells
hsa-miR-181a-5p27703791glioblast,
myeloid cells,
Embryonic stem
cells
hsa-miR-182-3p27763797immune cellscolonrectal cancer,immune
autoimmneresponse
hsa-miR-182-5p27783799lung, immuneautoimmuneimmune
cellsresponse
hsa-miR-197-3p28273848blood (myeloid),various cancers
other tissues(thyroid tumor,
leukemia, etc)
hsa-miR-197-5p28283849blood (myeloid),various cancers
other tissues(thyroid tumor,
leukemia, etc)
hsa-miR-21-3p28793099glioblast, Bloodautoimmune, heart
(meyloid cells),diseases, cancers
liver, vascular
endothelial cells
hsa-miR-214-3p28803901immune cells,varioua cancersimmune
pancreas(melanoma,response
pancreatic, ovarian)
hsa-miR-214-5p28813902immune cells,varioua cancersimmune
pancreas(melanoma,response
pancreatic, ovarian)
hsa-miR-21-5p28833904blood (myeloidautoimmune, heart
cells), liver,diseases, cancers
endothelial cells
hsa-miR-221-3p28943915endothelial cells,breastangiogenesis/
immune cellscancer, upregulatedvasculogenesis
in thyroid cell
transformation
induced by
HMGA1, TLR
signal pathway in
endotoxin tolerance,
upregulated in T cell
ALL
hsa-miR-221-5p28953916endothelialbreastangiogenesis/
cells, immunecancer, upregulatedvasculogenesis
cellsin thyroid cell
transformation
induced by
HMGA1, TLR
signal pathway in
endotoxin tolerance,
upregulated in T
cell ALL
hsa-miR-223-3p28983919meyloid cellsassociated with
CLL
hsa-miR-223-5p28993920meyloid cellsassociated with
CLL
hsa-miR-23b-3p29133934blood, myeloidcancers (renal
cellscancer,
glioblastoma,
prostate, etc)
and autoimmune
hsa-miR-23b-5p29143935blood, myeloidcancers(glioblastoma,
cellsprostate, etc) and
autoimmune
hsa-miR-24-1-5p29163937lung, myeloid
cells
hsa-miR-24-2-5p29173938lung, myeloid
cells
hsa-miR-24-3p29183939lung, myeloid
cells
hsa-miR-26a-1-29273948embryonic stemchronic lymphocytecell cycle and
3pcells, blood (Tleukemia and otherdifferentiation
cells)cancers
hsa-miR-26a-2-29283949blood (Tcells),chronic lymphocytecell cycle and
3pother tissuesleukemia and otherdifferentiation
cancers
hsa-miR-26a-5p29293950blood (Tcells),chronic lymphocytecell cycle and
other tissuesleukemia and otherdifferentiation
cancers
hsa-miR-26b-3p29303951hematopoietic
cells
hsa-miR-26b-5p29313952hematopoietic
cells
hsa-miR-27a-3p29323953myeloid cellsvarious cancer cells
hsa-miR-27a-5p29333954myeloid cellsvarious cancer cells
hsa-miR-27b-3p29343955myeloid cells,various cancer cellspro-angiogenic
vascular
endothelial cells
hsa-miR-28-3p29363957blood(immuneB/T cell lymphoma
cells)
hsa-miR-28-5p29373958blood(immuneB/T cell lymphoma
cells)
hsa-miR-290929393960T-Lymphocytes
hsa-miR-29a-3p29483969immuno system,various cancers,tumor
colonrectunneurodegenativesuppression,
diseaseimmune
modulation
(mir-29 family)
hsa-miR-29a-5p29493970immuno system,various cancers,adaptive
colonrectunneurodegenativeimmunity
disease
hsa-miR-29b-1-29503971immuno systemassociated withadaptive
5pCLL, other cancers,immunity
neurodegenative
disease
hsa-miR-29b-2-29513972immuno systemassociated withadaptive
5pCLL, other cancers,immunity
hsa-miR-29b-3p29523973immuno systemassociated withadaptive
CLL, other cancersimmunity
hsa-miR-29c-3p29533974immuno systemassociated withadaptive
CLL, other cancersimmunity
hsa-miR-29c-5p29543975immuno systemassociated withadaptive
CLL, other cancersimmunity
hsa-miR-30e-3p29844005myeloid cells,
glia cells
hsa-miR-30e-5p29854006myeloid cells,
glia cells
hsa-miR-331-5p31304151lymphocytes
hsa-miR-339-3p31374158immune cells
hsa-miR-339-5p31384159immune cells
hsa-miR-345-3p31474168hematopoieticincreased in
cellsfollicular
lymphoma(53),
other cancers
hsa-miR-345-5p31484169hematopoieticincreased in
cellsfollicular
lymphoma(53)
hsa-miR-34631494170immume cellscancers and
autoimmune
hsa-miR-34a-3p31504171breast, myeloidgastric cancer,tumor
cells, ciliatedCLL, othersuppressor, p53
epithelial cellsinducible
hsa-miR-34a-5p31514172breast, myeloidgastric cancer,tumor
cells, ciliatedCLL, othersuppressor, p53
epithelial cellsinducible
hsa-miR-363-3p31934214kidney stem cell,
blood cells
hsa-miR-363-5p31944215kidney stem cell,
blood cells
hsa-miR-37232774298hematopoietic
cells, lung,
placental (blood)
hsa-miR-377-3p32944315hematopoietic
cells
hsa-miR-377-5p32954316hematopoietic
cells
hsa-miR-493-3p49475968myeloid cells,
pancreas (islet)
hsa-miR-493-5p49485969myeloid cells,
pancreas (islet)
hsa-miR-542-3p51066127monocytestargets to
survivin,
introduce
growth arrest
hsa-miR-548b-51576178immune cells
5pfrontal cortex
hsa-miR-548c-5p51596180immune cells
frontal cortex
hsa-miR-548i51686189embryonic stem
cells (41),
immune cells
hsa-miR-548j51696190immune cells
hsa-miR-548n51736194embryonic stem
cells, immune
cells
hsa-miR-574-3p52796300blood (myeloidincreased in
cells)follicular
lymphoma(53)
hsa-miR-59853106331in blood
lymphocytes
(PBL)
hsa-miR-93555476568identified inassociated with
human cervicalenergy
cancermetabolism/obesity,
bloodmedullablastoma/
mononuclearneural stem cells
cells
hsa-miR-99a-3p55676588hemapoietic cells
hsa-miR-99a-5p55686589hemapoietic
cells, plasma
(exosome)
hsa-miR-99b-3p55696590hemapoietic
cells, Embryonic
stem cells,
hsa-miR-99b-5p55706591hemapoietic
cells, Embryonic
stem cells,
plasma(exosome)

III. MODIFICATIONS

Herein, in a signal-sensor polynucleotide (such as a primary construct or a mRNA molecule), the terms “modification” or, as appropriate, “modified” refer to modification with respect to A, G, U or C ribonucleotides. Generally, herein, these terms are not intended to refer to the ribonucleotide modifications in naturally occurring 5′-terminal mRNA cap moieties. In a polypeptide, the term “modification” refers to a modification as compared to the canonical set of 20 amino acids.

The modifications may be various distinct modifications. In some embodiments, the coding region, the flanking regions and/or the terminal regions may contain one, two, or more (optionally different) nucleoside or nucleotide modifications. In some embodiments, a modified signal-sensor polynucleotide, primary construct, or mmRNA introduced to a cell may exhibit reduced degradation in the cell, as compared to an unmodified signal-sensor polynucleotide, primary construct, or mmRNA.

The signal-sensor polynucleotides, primary constructs, and mmRNA can include any useful modification, such as to the sugar, the nucleobase, or the internucleoside linkage (e.g. to a linking phosphate/to a phosphodiester linkage/to the phosphodiester backbone). One or more atoms of a pyrimidine nucleobase may be replaced or substituted with optionally substituted amino, optionally substituted thiol, optionally substituted alkyl (e.g., methyl or ethyl), or halo (e.g., chloro or fluoro). In certain embodiments, modifications (e.g., one or more modifications) are present in each of the sugar and the internucleoside linkage. Modifications according to the present invention may be modifications of ribonucleic acids (RNAs) to deoxyribonucleic acids (DNAs), threose nucleic acids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs), locked nucleic acids (LNAs) or hybrids thereof). Additional modifications are described herein.

As described herein, in some embodiments, the signal-sensor polynucleotides, primary constructs, and mmRNA of the invention do not substantially induce an innate immune response of a cell into which the mRNA is introduced. Features of an induced innate immune response include 1) increased expression of pro-inflammatory cytokines, 2) activation of intracellular PRRs (RIG-I, MDA5, etc, and/or 3) termination or reduction in protein translation. In other embodiments, an immune response is induced.

In certain embodiments, it may desirable to intracellularly degrade a modified nucleic acid molecule introduced into the cell. For example, degradation of a modified nucleic acid molecule may be preferable if precise timing of protein production is desired. Thus, in some embodiments, the invention provides a modified nucleic acid molecule containing a degradation domain, which is capable of being acted on in a directed manner within a cell.

In another aspect, the present disclosure provides signal-sensor polynucleotides comprising a nucleoside or nucleotide that can disrupt the binding of a major groove interacting, e.g. binding, partner with the polynucleotide (e.g., where the modified nucleotide has decreased binding affinity to major groove interacting partner, as compared to an unmodified nucleotide).

The signal-sensor polynucleotides, primary constructs, and mmRNA can optionally include other agents (e.g., RNAi-inducing agents, RNAi agents, siRNAs, shRNAs, miRNAs, antisense RNAs, ribozymes, catalytic DNA, tRNA, RNAs that induce triple helix formation, aptamers, vectors, etc.). In some embodiments, the signal-sensor polynucleotides, primary constructs, or mmRNA may include one or more messenger RNAs (mRNAs) and one or more modified nucleoside or nucleotides (e.g., mmRNA molecules). Details for these signal-sensor polynucleotides, primary constructs, and mmRNA follow.

Signal-Sensor Polynucleotides and Primary Constructs

The signal-sensor polynucleotides, primary constructs, and mmRNA of the invention includes a first region of linked nucleosides encoding an oncology-related polypeptide of interest, a first flanking region located at the 5′ terminus of the first region, and a second flanking region located at the 3′ terminus of the first region.

In some embodiments, the signal-sensor polynucleotide, primary construct, or mmRNA are constructed according to the methods and modifications of International Application PCT/US12/058519 filed Oct. 3, 2012 (M9), the contents of which are incorporated herein by reference in their entirety.

The signal-sensor polynucleotides, primary constructs, and mmRNA can optionally include 5′ and/or 3′ flanking regions, which are described herein.

Signal-Sensor Modified RNA (mmRNA) Molecules

The present invention also includes the building blocks, e.g., modified ribonucleosides, modified ribonucleotides, of modified signal-sensor mRNA (mmRNA) molecules. For example, these building blocks can be useful for preparing the signal-sensor polynucleotides, primary constructs, or mmRNA of the invention. Such building blocks are taught in co-pending International Application PCT/US12/058519 filed Oct. 3, 2012 (M9), the contents of which are incorporated herein by reference in their entirety.

Modifications on the Nucleobase

The present disclosure provides for modified nucleosides and nucleotides. As described herein “nucleoside” is defined as a compound containing a sugar molecule (e.g., a pentose or ribose) or a derivative thereof in combination with an organic base (e.g., a purine or pyrimidine) or a derivative thereof (also referred to herein as “nucleobase”). As described herein, “nucleotide” is defined as a nucleoside including a phosphate group. In some embodiments, the nucleosides and nucleotides described herein are generally chemically modified on the major groove face. Exemplary non-limiting modifications include an amino group, a thiol group, an alkyl group, a halo group, or any described herein. The modified nucleotides may by synthesized by any useful method, as described herein (e.g., chemically, enzymatically, or recombinantly to include one or more modified or non-natural nucleosides).

The modified nucleosides and nucleotides can include a modified nucleobase. Examples of nucleobases found in RNA include, but are not limited to, adenine, guanine, cytosine, and uracil. Examples of nucleobase found in DNA include, but are not limited to, adenine, guanine, cytosine, and thymine. These nucleobases can be modified or wholly replaced to provide signal-sensor polynucleotides, primary constructs, or mmRNA molecules having enhanced properties. For example, the nucleosides and nucleotides described herein can be chemically modified. In some embodiments, chemical modifications can include an amino group, a thiol group, an alkyl group, or a halo group.

Modifications on the Internucleoside Linkage

The modified nucleotides, which may be incorporated into a signal-sensor polynucleotide, primary construct, or mmRNA molecule, can be modified on the internucleoside linkage (e.g., phosphate backbone). Herein, in the context of the polynucleotide backbone, the phrases “phosphate” and “phosphodiester” are used interchangeably. Backbone phosphate groups can be modified by replacing one or more of the oxygen atoms with a different substituent. Further, the modified nucleosides and nucleotides can include the wholesale replacement of an unmodified phosphate moiety with another internucleoside linkage as described herein. Examples of modified phosphate groups include, but are not limited to, phosphorothioate, phosphoroselenates, boranophosphates, boranophosphate esters, hydrogen phosphonates, phosphoramidates, phosphorodiamidates, alkyl or aryl phosphonates, and phosphotriesters. Phosphorodithioates have both non-linking oxygens replaced by sulfur. The phosphate linker can also be modified by the replacement of a linking oxygen with nitrogen (bridged phosphoramidates), sulfur (bridged phosphorothioates), and carbon (bridged methylene-phosphonates).

The α-thio substituted phosphate moiety is provided to confer stability to RNA and DNA polymers through the unnatural phosphorothioate backbone linkages. Phosphorothioate DNA and RNA have increased nuclease resistance and subsequently a longer half-life in a cellular environment. Phosphorothioate linked signal-sensor polynucleotides, primary constructs, or mmRNA molecules are expected to also reduce the innate immune response through weaker binding/activation of cellular innate immune molecules.

In specific embodiments, a modified nucleoside includes an alpha-thio-nucleoside (e.g., 5′-O-(1-thiophosphate)-adenosine, 5′-O-(1-thiophosphate)-cytidine (α-thio-cytidine), 5′-O-(1-thiophosphate)-guanosine, 5′-O-(1-thiophosphate)-uridine, or 5′-O-(1-thiophosphate)-pseudouridine).

Other internucleoside linkages that may be employed according to the present invention, including internucleoside linkages which do not contain a phosphorous atom, are described herein below.

Combinations of Modified Sugars, Nucleobases, and Internucleoside Linkages

The signal-sensor polynucleotides, primary constructs, and mmRNA of the invention can include a combination of modifications to the sugar, the nucleobase, and/or the internucleoside linkage. These combinations can include any one or more modifications described herein or in International Application PCT/US12/058519 filed Oct. 3, 2012 (M9), the contents of which are incorporated herein by reference in their entirety.

Synthesis of Signal-Sensor Primary Constructs, and mmRNA Molecules

The signal-sensor polypeptides, primary constructs, and mmRNA molecules for use in accordance with the invention may be prepared according to any useful technique, as described herein. The modified nucleosides and nucleotides used in the synthesis of signal-sensor polynucleotides, primary constructs, and mmRNA molecules disclosed herein can be prepared from readily available starting materials using the following general methods and procedures. Where typical or preferred process conditions (e.g., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are provided, a skilled artisan would be able to optimize and develop additional process conditions. Optimum reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization procedures.

The processes described herein can be monitored according to any suitable method known in the art. For example, product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1H or 13C) infrared spectroscopy, spectrophotometry (e.g., UV-visible), or mass spectrometry, or by chromatography such as high performance liquid chromatography (HPLC) or thin layer chromatography.

Preparation of signal-sensor polynucleotides, primary constructs, and mmRNA molecules of the present invention can involve the protection and deprotection of various chemical groups. The need for protection and deprotection, and the selection of appropriate protecting groups can be readily determined by one skilled in the art. The chemistry of protecting groups can be found, for example, in Greene, et al., Protective Groups in Organic Synthesis, 2d. Ed., Wiley & Sons, 1991, which is incorporated herein by reference in its entirety.

The reactions of the processes described herein can be carried out in suitable solvents, which can be readily selected by one of skill in the art of organic synthesis. Suitable solvents can be substantially nonreactive with the starting materials (reactants), the intermediates, or products at the temperatures at which the reactions are carried out, i.e., temperatures which can range from the solvent's freezing temperature to the solvent's boiling temperature. A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the particular reaction step, suitable solvents for a particular reaction step can be selected.

Resolution of racemic mixtures of modified nucleosides and nucleotides (e.g., mmRNA molecules) can be carried out by any of numerous methods known in the art. An example method includes fractional recrystallization using a “chiral resolving acid” which is an optically active, salt-forming organic acid. Suitable resolving agents for fractional recrystallization methods are, for example, optically active acids, such as the D and L forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid or the various optically active camphorsulfonic acids. Resolution of racemic mixtures can also be carried out by elution on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). Suitable elution solvent composition can be determined by one skilled in the art.

Modified nucleosides and nucleotides (e.g., building block molecules) can be prepared according to the synthetic methods described in Ogata et al., J. Org. Chem. 74:2585-2588 (2009); Purmal et al., Nucl. Acids Res. 22(1): 72-78, (1994); Fukuhara et al., Biochemistry, 1(4): 563-568 (1962); and Xu et al., Tetrahedron, 48(9): 1729-1740 (1992), each of which are incorporated by reference in their entirety.

The signal-sensor polynucleotides, primary constructs, and mmRNA of the invention may or may not be uniformly modified along the entire length of the molecule. For example, one or more or all types of nucleotide (e.g., purine or pyrimidine, or any one or more or all of A, G, U, C) may or may not be uniformly modified in a polynucleotide of the invention, or in a given predetermined sequence region thereof (e.g. one or more of the sequence regions represented in FIG. 1). In some embodiments, all nucleotides X in a signal-sensor polynucleotide of the invention (or in a given sequence region thereof) are modified, wherein X may any one of nucleotides A, G, U, C, or any one of the combinations A+G, A+U, A+C, G+U, G+C, U+C, A+G+U, A+G+C, G+U+C or A+G+C.

Different sugar modifications, nucleotide modifications, and/or internucleoside linkages (e.g., backbone structures) may exist at various positions in the signal-sensor polynucleotide, primary construct, or mmRNA. One of ordinary skill in the art will appreciate that the nucleotide analogs or other modification(s) may be located at any position(s) of a signal-sensor polynucleotide, primary construct, or mmRNA such that the function of the signal-sensor polynucleotide, primary construct, or mmRNA is not substantially decreased. A modification may also be a 5′ or 3′ terminal modification. The signal-sensor polynucleotide, primary construct, or mmRNA may contain from about 1% to about 100% modified nucleotides (either in relation to overall nucleotide content, or in relation to one or more types of nucleotide, i.e. any one or more of A, G, U or C) or any intervening percentage (e.g., from 1% to 20%, from 1% to 25%, from 1% to 50%, from 1% to 60%, from 1% to 70%, from 1% to 80%, from 1% to 90%, from 1% to 95%, from 10% to 20%, from 10% to 25%, from 10% to 50%, from 10% to 60%, from 10% to 70%, from 10% to 80%, from 10% to 90%, from 10% to 95%, from 10% to 100%, from 20% to 25%, from 20% to 50%, from 20% to 60%, from 20% to 70%, from 20% to 80%, from 20% to 90%, from 20% to 95%, from 20% to 100%, from 50% to 60%, from 50% to 70%, from 50% to 80%, from 50% to 90%, from 50% to 95%, from 50% to 100%, from 70% to 80%, from 70% to 90%, from 70% to 95%, from 70% to 100%, from 80% to 90%, from 80% to 95%, from 80% to 100%, from 90% to 95%, from 90% to 100%, and from 95% to 100%).

In some embodiments, the signal-sensor polynucleotide, primary construct, or mmRNA includes a modified pyrimidine (e.g., a modified uracil/uridine/U or modified cytosine/cytidine/C). In some embodiments, the uracil or uridine (generally: U) in the signal-sensor polynucleotide, primary construct, or mmRNA molecule may be replaced with from about 1% to about 100% of a modified uracil or modified uridine (e.g., from 1% to 20%, from 1% to 25%, from 1% to 50%, from 1% to 60%, from 1% to 70%, from 1% to 80%, from 1% to 90%, from 1% to 95%, from 10% to 20%, from 10% to 25%, from 10% to 50%, from 10% to 60%, from 10% to 70%, from 10% to 80%, from 10% to 90%, from 10% to 95%, from 10% to 100%, from 20% to 25%, from 20% to 50%, from 20% to 60%, from 20% to 70%, from 20% to 80%, from 20% to 90%, from 20% to 95%, from 20% to 100%, from 50% to 60%, from 50% to 70%, from 50% to 80%, from 50% to 90%, from 50% to 95%, from 50% to 100%, from 70% to 80%, from 70% to 90%, from 70% to 95%, from 70% to 100%, from 80% to 90%, from 80% to 95%, from 80% to 100%, from 90% to 95%, from 90% to 100%, and from 95% to 100% of a modified uracil or modified uridine). The modified uracil or uridine can be replaced by a compound having a single unique structure or by a plurality of compounds having different structures (e.g., 2, 3, 4 or more unique structures, as described herein). In some embodiments, the cytosine or cytidine (generally: C) in the signal-sensor polynucleotide, primary construct, or mmRNA molecule may be replaced with from about 1% to about 100% of a modified cytosine or modified cytidine (e.g., from 1% to 20%, from 1% to 25%, from 1% to 50%, from 1% to 60%, from 1% to 70%, from 1% to 80%, from 1% to 90%, from 1% to 95%, from 10% to 20%, from 10% to 25%, from 10% to 50%, from 10% to 60%, from 10% to 70%, from 10% to 80%, from 10% to 90%, from 10% to 95%, from 10% to 100%, from 20% to 25%, from 20% to 50%, from 20% to 60%, from 20% to 70%, from 20% to 80%, from 20% to 90%, from 20% to 95%, from 20% to 100%, from 50% to 60%, from 50% to 70%, from 50% to 80%, from 50% to 90%, from 50% to 95%, from 50% to 100%, from 70% to 80%, from 70% to 90%, from 70% to 95%, from 70% to 100%, from 80% to 90%, from 80% to 95%, from 80% to 100%, from 90% to 95%, from 90% to 100%, and from 95% to 100% of a modified cytosine or modified cytidine). The modified cytosine or cytidine can be replaced by a compound having a single unique structure or by a plurality of compounds having different structures (e.g., 2, 3, 4 or more unique structures, as described herein).

Combinations of Nucleotides

Further examples of modified nucleotides and modified nucleotide combinations are provided in International Application PCT/US12/058519 filed Oct. 3, 2012 (M9) the contents of which are incorporated herein by reference in their entirety.

In some embodiments, at least 25% of the cytidines are replaced (e.g., at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100%).

In some embodiments, at least 25% of the uracils are replaced (e.g., at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100%).

In some embodiments, at least 25% of the cytidines are replaced, and at least 25% of the uracils are replaced (e.g., at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100%).

IV. PHARMACEUTICAL COMPOSITIONS

Formulation, Administration, Delivery and Dosing

The present invention provides signal-sensor polynucleotides, primary constructs and mmRNA compositions and complexes in combination with one or more pharmaceutically acceptable excipients. Pharmaceutical compositions may optionally comprise one or more additional active substances, e.g. therapeutically and/or prophylactically active substances. General considerations in the formulation and/or manufacture of pharmaceutical agents may be found, for example, in Remington: The Science and Practice of Pharmacy 21st ed., Lippincott Williams & Wilkins, 2005 (incorporated herein by reference).

In some embodiments, compositions are administered to humans, human patients or subjects. For the purposes of the present disclosure, the phrase “active ingredient” generally refers to signal-sensor polynucleotides, primary constructs and mmRNA to be delivered as described herein.

Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to any other animal, e.g., to non-human animals, e.g. non-human mammals. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with merely ordinary, if any, experimentation. Subjects to which administration of the pharmaceutical compositions is contemplated include, but are not limited to, humans and/or other primates; mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, dogs, mice, and/or rats; and/or birds, including commercially relevant birds such as poultry, chickens, ducks, geese, and/or turkeys.

Formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with an excipient and/or one or more other accessory ingredients, and then, if necessary and/or desirable, dividing, shaping and/or packaging the product into a desired single- or multi-dose unit.

A pharmaceutical composition in accordance with the invention may be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. As used herein, a “unit dose” is discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.

Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition in accordance with the invention will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered. By way of example, the composition may comprise between 0.1% and 100%, e.g., between 0.5 and 50%, between 1-30%, between 5-80%, at least 80% (w/w) active ingredient.

Formulations

The signal-sensor polynucleotide, primary construct, and mmRNA of the invention can be formulated using one or more excipients to: (1) increase stability; (2) increase cell transfection; (3) permit the sustained or delayed release (e.g., from a depot formulation of the signal-sensor polynucleotide, primary construct, or mmRNA); (4) alter the biodistribution (e.g., target the polynucleotide, primary construct, or mmRNA to specific tissues or cell types); (5) increase the translation of encoded protein in vivo; and/or (6) alter the release profile of encoded protein in vivo. In addition to traditional excipients such as any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, excipients of the present invention can include, without limitation, lipidoids, liposomes, lipid nanoparticles, polymers, lipoplexes, core-shell nanoparticles, peptides, proteins, cells transfected with signal-sensor polynucleotide, primary construct, or mmRNA (e.g., for transplantation into a subject), hyaluronidase, nanoparticle mimics and combinations thereof. Further, the signal-sensor polynucleotide, primary construct, or mmRNA of the present invention may be formulated using self-assembled nucleic acid nanoparticles.

Accordingly, the formulations of the invention can include one or more excipients, each in an amount that together increases the stability of the signal-sensor polynucleotide, primary construct, or mmRNA, increases cell transfection by the signal-sensor polynucleotide, primary construct, or mmRNA, increases the expression of polynucleotide, primary construct, or mmRNA encoded protein, and/or alters the release profile of signal-sensor polynucleotide, primary construct, or mmRNA encoded proteins. Further, the primary construct and mmRNA of the present invention may be formulated using self-assembled nucleic acid nanoparticles.

Formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of associating the active ingredient with an excipient and/or one or more other accessory ingredients.

A pharmaceutical composition in accordance with the present disclosure may be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. As used herein, a “unit dose” refers to a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient may generally be equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage including, but not limited to, one-half or one-third of such a dosage.

Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition in accordance with the present disclosure may vary, depending upon the identity, size, and/or condition of the subject being treated and further depending upon the route by which the composition is to be administered. For example, the composition may comprise between 0.1% and 99% (w/w) of the active ingredient.

In some embodiments, the formulations described herein may contain at least one signal-sensor mmRNA. As a non-limiting example, the formulations may contain 1, 2, 3, 4 or 5 signal-sensor mmRNA. In one embodiment the formulation may contain modified mRNA encoding proteins selected from categories such as, proteins. In one embodiment, the formulation contains at least three signal-sensor modified mRNA encoding oncology-related proteins. In one embodiment, the formulation contains at least five signal-sensor modified mRNA encoding oncology-related proteins.

Pharmaceutical formulations may additionally comprise a pharmaceutically acceptable excipient, which, as used herein, includes, but is not limited to, any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, and the like, as suited to the particular dosage form desired. Various excipients for formulating pharmaceutical compositions and techniques for preparing the composition are known in the art (see Remington: The Science and Practice of Pharmacy, 21st Edition, A. R. Gennaro, Lippincott, Williams & Wilkins, Baltimore, Md., 2006; incorporated herein by reference). The use of a conventional excipient medium may be contemplated within the scope of the present disclosure, except insofar as any conventional excipient medium may be incompatible with a substance or its derivatives, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutical composition.

In some embodiments, the particle size of the lipid nanoparticle may be increased and/or decreased. The change in particle size may be able to help counter biological reaction such as, but not limited to, inflammation or may increase the biological effect of the signal-sensor modified mRNA delivered to mammals.

Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, surface active agents and/or emulsifiers, preservatives, buffering agents, lubricating agents, and/or oils. Such excipients may optionally be included in the pharmaceutical formulations of the invention.

Pharmaceutical compositions of the present invention may comprise at least one adjuvant which may be a chemo-adjuvant. Non-limiting examples of chemo-adjuvants and delivery systems which comprises a chemo-adjuvant are described in International Patent Publication No. WO2013134349, the contents of which is herein incorporated by reference in its entirety. The chemo-adjuvant may be bonded to, non-covalently bonded to or encapsulated within a delivery vehicle described herein.

Lipidoids

The synthesis of lipidoids has been extensively described and formulations containing these compounds are particularly suited for delivery of signal-sensor polynucleotides, primary constructs or mmRNA (see Mahon et al., Bioconjug Chem. 2010 21:1448-1454; Schroeder et al., J Intern Med. 2010 267:9-21; Akinc et al., Nat Biotechnol. 2008 26:561-569; Love et al., Proc Natl Acad Sci USA. 2010 107:1864-1869; Siegwart et al., Proc Natl Acad Sci USA. 2011 108:12996-3001; all of which are incorporated herein in their entireties).

While these lipidoids have been used to effectively deliver double stranded small interfering RNA molecules in rodents and non-human primates (see Akinc et al., Nat Biotechnol. 2008 26:561-569; Frank-Kamenetsky et al., Proc Natl Acad Sci USA. 2008 105:11915-11920; Akinc et al., Mol Ther. 2009 17:872-879; Love et al., Proc Natl Acad Sci USA. 2010 107:1864-1869; Leuschner et al., Nat Biotechnol. 2011 29:1005-1010; all of which is incorporated herein in their entirety), the present disclosure describes their formulation and use in delivering single stranded signal-sensor polynucleotides, primary constructs, or mmRNA. Complexes, micelles, liposomes or particles can be prepared containing these lipidoids and therefore, can result in an effective delivery of the signal-sensor polynucleotide, primary construct, or mmRNA, as judged by the production of an encoded protein, following the injection of a lipidoid formulation via localized and/or systemic routes of administration. Lipidoid complexes of signal-sensor polynucleotides, primary constructs, or mmRNA can be administered by various means including, but not limited to, intravenous, intramuscular, or subcutaneous routes.

In vivo delivery of nucleic acids may be affected by many parameters, including, but not limited to, the formulation composition, nature of particle PEGylation, degree of loading, oligonucleotide to lipid ratio, and biophysical parameters such as particle size (Akinc et al., Mol Ther. 2009 17:872-879; herein incorporated by reference in its entirety). As an example, small changes in the anchor chain length of poly(ethylene glycol) (PEG) lipids may result in significant effects on in vivo efficacy. Formulations with the different lipidoids, including, but not limited to penta[3-(1-laurylaminopropionyl)]-triethylenetetramine hydrochloride (TETA-5LAP; aka 98N12-5, see Murugaiah et al., Analytical Biochemistry, 401:61 (2010)), C12-200 (including derivatives and variants), and MD1, can be tested for in vivo activity.

The lipidoid referred to herein as “98N12-5” is disclosed by Akinc et al., Mol Ther. 2009 17:872-879 and is incorporated by reference in its entirety.

The lipidoid referred to herein as “C12-200” is disclosed by Love et al., Proc Natl Acad Sci USA. 2010 107:1864-1869 and Liu and Huang, Molecular Therapy. 2010 669-670; both of which are herein incorporated by reference in their entirety. The lipidoid formulations can include particles comprising either 3 or 4 or more components in addition to signal-sensor polynucleotide, primary construct, or mmRNA. As an example, formulations with certain lipidoids, include, but are not limited to, 98N12-5 and may contain 42% lipidoid, 48% cholesterol and 10% PEG (C14 alkyl chain length). As another example, formulations with certain lipidoids, include, but are not limited to, C12-200 and may contain 50% lipidoid, 10% disteroylphosphatidyl choline, 38.5% cholesterol, and 1.5% PEG-DMG.

Combinations of different lipidoids may be used to improve the efficacy of signal-sensor polynucleotide, primary construct, or mmRNA directed protein production as the lipidoids may be able to increase cell transfection by the signal-sensor polynucleotide, primary construct, or mmRNA; and/or increase the translation of encoded oncology-related protein (see Whitehead et al., Mol. Ther. 2011, 19:1688-1694, herein incorporated by reference in its entirety).

In some embodiments, the particle size of the lipid nanoparticle may be increased and/or decreased. The change in particle size may be able to help counter biological reaction such as, but not limited to, inflammation or may increase the biological effect of, the signal-sensor polynucleotide, primary construct, or mmRNA delivered to subjects.

Liposomes, Lipoplexes, and Lipid Nanoparticles

The signal-sensor polynucleotide, primary construct, and mmRNA of the invention can be formulated using one or more liposomes, lipoplexes, or lipid nanoparticles. In one embodiment, pharmaceutical compositions of signal-sensor polynucleotide, primary construct, or mmRNA include liposomes. Liposomes are artificially-prepared vesicles which may primarily be composed of a lipid bilayer and may be used as a delivery vehicle for the administration of nutrients and pharmaceutical formulations. Liposomes can be of different sizes such as, but not limited to, a multilamellar vesicle (MLV) which may be hundreds of nanometers in diameter and may contain a series of concentric bilayers separated by narrow aqueous compartments, a small unicellular vesicle (SUV) which may be smaller than 50 nm in diameter, and a large unilamellar vesicle (LUV) which may be between 50 and 500 nm in diameter. Liposome design may include, but is not limited to, opsonins or ligands in order to improve the attachment of liposomes to unhealthy tissue or to activate events such as, but not limited to, endocytosis. Liposomes may contain a low or a high pH in order to improve the delivery of the pharmaceutical formulations.

The formation of liposomes may depend on the physicochemical characteristics such as, but not limited to, the pharmaceutical formulation entrapped and the liposomal ingredients, the nature of the medium in which the lipid vesicles are dispersed, the effective concentration of the entrapped substance and its potential toxicity, any additional processes involved during the application and/or delivery of the vesicles, the optimization size, polydispersity and the shelf-life of the vesicles for the intended application, and the batch-to-batch reproducibility and possibility of large-scale production of safe and efficient liposomal products.

In one embodiment, pharmaceutical compositions described herein may include, without limitation, liposomes such as those formed from 1,2-dioleyloxy-N,N-dimethylaminopropane (DODMA) liposomes, DiLa2 liposomes from Marina Biotech (Bothell, Wash.), 1,2-dilinoleyloxy-3-dimethylaminopropane (DLin-DMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLin-KC2-DMA), and MC3 (US20100324120; herein incorporated by reference in its entirety) and liposomes which may deliver small molecule drugs such as, but not limited to, DOXIL® from Janssen Biotech, Inc. (Horsham, Pa.).

In one embodiment, pharmaceutical compositions described herein may include, without limitation, liposomes such as those formed from the synthesis of stabilized plasmid-lipid particles (SPLP) or stabilized nucleic acid lipid particle (SNALP) that have been previously described and shown to be suitable for oligonucleotide delivery in vitro and in vivo (see Wheeler et al. Gene Therapy. 1999 6:271-281; Zhang et al. Gene Therapy. 1999 6:1438-1447; Jeffs et al. Pharm Res. 2005 22:362-372; Morrissey et al., Nat Biotechnol. 2005 2:1002-1007; Zimmermann et al., Nature. 2006 441:111-114; Heyes et al. J Contr Rel. 2005 107:276-287; Semple et al. Nature Biotech. 2010 28:172-176; Judge et al. J Clin Invest. 2009 119:661-673; deFougerolles Hum Gene Ther. 2008 19:125-132; all of which are incorporated herein in their entireties.) The original manufacture method by Wheeler et al. was a detergent dialysis method, which was later improved by Jeffs et al. and is referred to as the spontaneous vesicle formation method. The liposome formulations are composed of 3 to 4 lipid components in addition to the signal-sensor polynucleotide, primary construct, or mmRNA. As an example a liposome can contain, but is not limited to, 55% cholesterol, 20% disteroylphosphatidyl choline (DSPC), 10% PEG-S-DSG, and 15% 1,2-dioleyloxy-N,N-dimethylaminopropane (DODMA), as described by Jeffs et al. As another example, certain liposome formulations may contain, but are not limited to, 48% cholesterol, 20% DSPC, 2% PEG-c-DMA, and 30% cationic lipid, where the cationic lipid can be 1,2-distearloxy-N,N-dimethylaminopropane (DSDMA), DODMA, DLin-DMA, or 1,2-dilinolenyloxy-3-dimethylaminopropane (DLenDMA), as described by Heyes et al.

In one embodiment, pharmaceutical compositions may include liposomes which may be formed to deliver signal-sensor mmRNA which may encode at least one immunogen. The mmRNA may be encapsulated by the liposome and/or it may be contained in an aqueous core which may then be encapsulated by the liposome (see International Pub. Nos. WO2012031046, WO2012031043, WO201203091 and WO2012006378 herein incorporated by reference in their entireties). In another embodiment, the signal-sensor mmRNA which may encode an immunogen may be formulated in a cationic oil-in-water emulsion where the emulsion particle comprises an oil core and a cationic lipid which can interact with the signal-sensor mmRNA anchoring the molecule to the emulsion particle (see International Pub. No. WO2012006380). In yet another embodiment, the lipid formulation may include at least cationic lipid, a lipid which may enhance transfection and a least one lipid which contains a hydrophilic head group linked to a lipid moiety (International Pub. No. WO2011076807 and U.S. Pub. No. 20110200582; herein incorporated by reference in their entireties). In another embodiment, the signal-sensor polynucleotides, primary constructs and/or mmRNA encoding an immunogen may be formulated in a lipid vesicle which may have crosslinks between functionalized lipid bilayers (see U.S. Pub. No. 20120177724, herein incorporated by reference in its entirety).

In one embodiment, the signal-sensor polynucleotides, primary constructs and/or mmRNA may be formulated in a lipid vesicle which may have crosslinks between functionalized lipid bilayers.

In one embodiment, the signal-sensor polynucleotides, primary constructs and/or mmRNA may be formulated in a lipid-polycation complex. The formation of the lipid-polycation complex may be accomplished by methods known in the art and/or as described in U.S. Pub. No. 20120178702, herein incorporated by reference in its entirety. As a non-limiting example, the polycation may include a cationic peptide or a polypeptide such as, but not limited to, polylysine, polyornithine and/or polyarginine. In another embodiment, the signal-sensor polynucleotides, primary constructs and/or mmRNA may be formulated in a lipid-polycation complex which may further include a neutral lipid such as, but not limited to, cholesterol or dioleoyl phosphatidylethanolamine (DOPE).

The liposome formulation may be influenced by, but not limited to, the selection of the cationic lipid component, the degree of cationic lipid saturation, the nature of the PEGylation, ratio of all components and biophysical parameters such as size. In one example by Semple et al. (Semple et al. Nature Biotech. 2010 28:172-176), the liposome formulation was composed of 57.1% cationic lipid, 7.1% dipalmitoylphosphatidylcholine, 34.3% cholesterol, and 1.4% PEG-c-DMA. As another example, changing the composition of the cationic lipid could more effectively deliver siRNA to various antigen presenting cells (Basha et al. Mol Ther. 2011 19:2186-2200; herein incorporated by reference in its entirety).

In some embodiments, the ratio of PEG in the LNP formulations may be increased or decreased and/or the carbon chain length of the PEG lipid may be modified from C14 to C18 to alter the pharmacokinetics and/or biodistribution of the LNP formulations. As a non-limiting example, LNP formulations may contain 1-5% of the lipid molar ratio of PEG-c-DOMG as compared to the cationic lipid, DSPC and cholesterol. In another embodiment the PEG-c-DOMG may be replaced with a PEG lipid such as, but not limited to, PEG-DSG (1,2-Distearoyl-sn-glycerol, methoxypolyethylene glycol) or PEG-DPG (1,2-Dipalmitoyl-sn-glycerol, methoxypolyethylene glycol). The cationic lipid may be selected from any lipid known in the art such as, but not limited to, DLin-MC3-DMA, DLin-DMA, C12-200 and DLin-KC2-DMA.

In one embodiment, the LNP formulations of the signal-sensor polynucleotides, primary constructs and/or mmRNA may contain PEG-c-DOMG 3% lipid molar ratio. In another embodiment, the LNP formulations of the signal-sensor polynucleotides, primary constructs and/or mmRNA may contain PEG-c-DOMG 1.5% lipid molar ratio.

In one embodiment, the pharmaceutical compositions of the signal-sensor polynucleotides, primary constructs and/or mmRNA may include at least one of the PEGylated lipids described in International Publication No. 2012099755, herein incorporated by reference.

In one embodiment, the pharmaceutical compositions may be formulated in liposomes such as, but not limited to, DiLa2 liposomes (Marina Biotech, Bothell, Wash.), SMARTICLES® (Marina Biotech, Bothell, Wash.), neutral DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine) based liposomes (e.g., siRNA delivery for ovarian cancer (Landen et al. Cancer Biology & Therapy 2006 5(12)1708-1713)) and hyaluronan-coated liposomes (Quiet Therapeutics, Israel).

In some embodiments the liposome may be a liposomal nanostructure which has been formulated for treatment of cancers and other diseases or to control the cholesterol metabolism in cells. The liposome nanostructure may also comprise a scavenger receptor type B-1 (SR-B1) in order to kill cancer cells. Non-limiting examples of liposomal nanostructures, which may be used with the signal-sensor polynucleotides described herein, are described in International Publication No. WO2013126776, the contents of which are herein incorporated by reference in its entirety.

In one embodiment, the liposomes described herein may comprise at least one immunomodulator such as, but not limited to, cytokines Formulations and methods of using the liposomes comprising at least one immunomodulator are described in International Publication No WO2013129935 and WO2013129936, the contents of each of which are herein incorporated by reference in their entirety. As a non-limiting example, the liposomes comprising at least one immunomodulator may be used in the treatment of cancer. The liposomes comprising an immunomodulator may comprise a signal-sensor polynucleotide described herein. As a non-limiting example, the liposome comprising an immunomodulator may be used in a combination with at least one antibody such as the particulate or vesicular immunomodulators described in International Publication No WO2013129936, the contents of which are herein incorporated by reference in its entirety.

Lipid nanoparticle formulations may be improved by replacing the cationic lipid with a biodegradable cationic lipid which is known as a rapidly eliminated lipid nanoparticle (reLNP). Ionizable cationic lipids, such as, but not limited to, DLinDMA, DLin-KC2-DMA, and DLin-MC3-DMA, have been shown to accumulate in plasma and tissues over time and may be a potential source of toxicity. The rapid metabolism of the rapidly eliminated lipids can improve the tolerability and therapeutic index of the lipid nanoparticles by an order of magnitude from a 1 mg/kg dose to a 10 mg/kg dose in rat. Inclusion of an enzymatically degraded ester linkage can improve the degradation and metabolism profile of the cationic component, while still maintaining the activity of the reLNP formulation. The ester linkage can be internally located within the lipid chain or it may be terminally located at the terminal end of the lipid chain. The internal ester linkage may replace any carbon in the lipid chain.

In one embodiment, the internal ester linkage may be located on either side of the saturated carbon.

In one embodiment, an immune response may be elicited by delivering a lipid nanoparticle which may include a nanospecies, a polymer and an immunogen. (U.S. Publication No. 20120189700 and International Publication No. WO2012099805; herein incorporated by reference in their entireties). The polymer may encapsulate the nanospecies or partially encapsulate the nanospecies. The immunogen may be a recombinant oncology-related protein, a signal-sensor modified RNA and/or a primary construct described herein. In one embodiment, the lipid nanoparticle may be formulated for use in a vaccine such as, but not limited to, against a pathogen.

Lipid nanoparticles may be engineered to alter the surface properties of particles so the lipid nanoparticles may penetrate the mucosal barrier. Mucus is located on mucosal tissue such as, but not limited to, oral (e.g., the buccal and esophageal membranes and tonsil tissue), ophthalmic, gastrointestinal (e.g., stomach, small intestine, large intestine, colon, rectum), nasal, respiratory (e.g., nasal, pharyngeal, tracheal and bronchial membranes), genital (e.g., vaginal, cervical and urethral membranes). Nanoparticles larger than 10-200 nm which are preferred for higher drug encapsulation efficiency and the ability to provide the sustained delivery of a wide array of drugs have been thought to be too large to rapidly diffuse through mucosal barriers. Mucus is continuously secreted, shed, discarded or digested and recycled so most of the trapped particles may be removed from the mucosla tissue within seconds or within a few hours. Large polymeric nanoparticles (200 nm-500 nm in diameter) which have been coated densely with a low molecular weight polyethylene glycol (PEG) diffused through mucus only 4 to 6-fold lower than the same particles diffusing in water (Lai et al. PNAS 2007 104(5):1482-487; Lai et al. Adv Drug Deliv Rev. 2009 61(2): 158-171; herein incorporated by reference in their entirety). The transport of nanoparticles may be determined using rates of permeation and/or fluorescent microscopy techniques including, but not limited to, fluorescence recovery after photobleaching (FRAP) and high resolution multiple particle tracking (MPT).

The lipid nanoparticle engineered to penetrate mucus may comprise a polymeric material (i.e. a polymeric core) and/or a polymer-vitamin conjugate and/or a tri-block co-polymer. The polymeric material may include, but is not limited to, polyamines, polyethers, polyamides, polyesters, polycarbamates, polyureas, polycarbonates, poly(styrenes), polyimides, polysulfones, polyurethanes, polyacetylenes, polyethylenes, polyethyeneimines, polyisocyanates, polyacrylates, polymethacrylates, polyacrylonitriles, and polyarylates. The polymeric material may be biodegradable and/or biocompatible. Non-limiting examples of specific polymers include poly(caprolactone) (PCL), ethylene vinyl acetate polymer (EVA), poly(lactic acid) (PLA), poly(L-lactic acid) (PLLA), poly(glycolic acid) (PGA), poly(lactic acid-co-glycolic acid) (PLGA), poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide) (PDLA), poly(L-lactide) (PLLA), poly(D,L-lactide-co-caprolactone), poly(D,L-lactide-co-caprolactone-co-glycolide), poly(D,L-lactide-co-PEO-co-D,L-lactide), poly(D,L-lactide-co-PPO-co-D,L-lactide), polyalkyl cyanoacralate, polyurethane, poly-L-lysine (PLL), hydroxypropyl methacrylate (HPMA), polyethyleneglycol, poly-L-glutamic acid, poly(hydroxy acids), polyanhydrides, polyorthoesters, poly(ester amides), polyamides, poly(ester ethers), polycarbonates, polyalkylenes such as polyethylene and polypropylene, polyalkylene glycols such as poly(ethylene glycol) (PEG), polyalkylene oxides (PEO), polyalkylene terephthalates such as poly(ethylene terephthalate), polyvinyl alcohols (PVA), polyvinyl ethers, polyvinyl esters such as poly(vinyl acetate), polyvinyl halides such as poly(vinyl chloride) (PVC), polyvinylpyrrolidone, polysiloxanes, polystyrene (PS), polyurethanes, derivatized celluloses such as alkyl celluloses, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitro celluloses, hydroxypropylcellulose, carboxymethylcellulose, polymers of acrylic acids, such as poly(methyl(meth)acrylate) (PMMA), poly(ethyl(meth)acrylate), poly(butyl(meth)acrylate), poly(isobutyl(meth)acrylate), poly(hexyl(meth)acrylate), poly(isodecyl(meth)acrylate), poly(lauryl(meth)acrylate), poly(phenyl(meth)acrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) and copolymers and mixtures thereof, polydioxanone and its copolymers, polyhydroxyalkanoates, polypropylene fumarate, polyoxymethylene, poloxamers, poly(ortho)esters, poly(butyric acid), poly(valeric acid), poly(lactide-co-caprolactone), and trimethylene carbonate, polyvinylpyrrolidone. The lipid nanoparticle may be coated or associated with a co-polymer such as, but not limited to, a block co-polymer, and (poly(ethylene glycol))-(poly(propylene oxide))-(poly(ethylene glycol))triblock copolymer (see US Publication 20120121718 and US Publication 20100003337; herein incorporated by reference in their entireties). The co-polymer may be a polymer that is generally regarded as safe (GRAS) and the formation of the lipid nanoparticle may be in such a way that no new chemical entities are created. For example, the lipid nanoparticle may comprise poloxamers coating PLGA nanoparticles without forming new chemical entities which are still able to rapidly penetrate human mucus (Yang et al. Angew. Chem. Int. Ed. 2011 50:2597-2600; herein incorporated by reference in its entirety).

The vitamin of the polymer-vitamin conjugate may be vitamin E. The vitamin portion of the conjugate may be substituted with other suitable components such as, but not limited to, vitamin A, vitamin E, other vitamins, cholesterol, a hydrophobic moiety, or a hydrophobic component of other surfactants (e.g., sterol chains, fatty acids, hydrocarbon chains and alkylene oxide chains).

The lipid nanoparticle engineered to penetrate mucus may include surface altering agents such as, but not limited to, signal-sensor mmRNA, anionic protein (e.g., bovine serum albumin), surfactants (e.g., cationic surfactants such as for example dimethyldioctadecyl-ammonium bromide), sugars or sugar derivatives (e.g., cyclodextrin), nucleic acids, polymers (e.g., heparin, polyethylene glycol and poloxamer), mucolytic agents (e.g., N-acetylcysteine, mugwort, bromelain, papain, clerodendrum, acetylcysteine, bromhexine, carbocisteine, eprazinone, mesna, ambroxol, sobrerol, domiodol, letosteine, stepronin, tiopronin, gelsolin, thymosin (34 dornase alfa, neltenexine, erdosteine) and various DNases including rhDNase. The surface altering agent may be embedded or enmeshed in the particle's surface or disposed (e.g., by coating, adsorption, covalent linkage, or other process) on the surface of the lipid nanoparticle. (see US Publication 20100215580 and US Publication 20080166414; herein incorporated by reference in their entireties).

The mucus penetrating lipid nanoparticles may comprise at least one signal-sensor mmRNA described herein. The signal-sensor mmRNA may be encapsulated in the lipid nanoparticle and/or disposed on the surface of the particle. The signal-sensor mmRNA may be covalently coupled to the lipid nanoparticle. Formulations of mucus penetrating lipid nanoparticles may comprise a plurality of nanoparticles. Further, the formulations may contain particles which may interact with the mucus and alter the structural and/or adhesive properties of the surrounding mucus to decrease mucoadhesion which may increase the delivery of the mucus penetrating lipid nanoparticles to the mucosal tissue.

Lipid nanoparticles may be engineered to alter the surface properties of particles so the lipid nanoparticles may penetrate the mucosal barrier. Mucus is located on mucosal tissue such as, but not limited to, oral (e.g., the buccal and esophageal membranes and tonsil tissue), ophthalmic, gastrointestinal (e.g., stomach, small intestine, large intestine, colon, rectum), nasal, respiratory (e.g., nasal, pharyngeal, tracheal and bronchial membranes), genital (e.g., vaginal, cervical and urethral membranes). Nanoparticles larger than 10-200 nm which are preferred for higher drug encapsulation efficiency and the ability to provide the sustained delivery of a wide array of drugs have been thought to be too large to rapidly diffuse through mucosal barriers. Mucus is continuously secreted, shed, discarded or digested and recycled so most of the trapped particles may be removed from the mucosla tissue within seconds or within a few hours. Large polymeric nanoparticles (200 nm-500 nm in diameter) which have been coated densely with a low molecular weight polyethylene glycol (PEG) diffused through mucus only 4 to 6-fold lower than the same particles diffusing in water (Lai et al. PNAS 2007 104(5):1482-487; Lai et al. Adv Drug Deliv Rev. 2009 61(2): 158-171; herein incorporated by reference in their entirety). The transport of nanoparticles may be determined using rates of permeation and/or fluorescent microscopy techniques including, but not limited to, fluorescence recovery after photobleaching (FRAP) and high resolution multiple particle tracking (MPT).

The lipid nanoparticle engineered to penetrate mucus may comprise a polymeric material (i.e. a polymeric core) and/or a polymer-vitamin conjugate and/or a tri-block co-polymer. The polymeric material may including, but is not limited to, polyamines, polyethers, polyamides, polyesters, polycarbamates, polyureas, polycarbonates, poly(styrenes), polyimides, polysulfones, polyurethanes, polyacetylenes, polyethylenes, polyethyeneimines, polyisocyanates, polyacrylates, polymethacrylates, polyacrylonitriles, and polyarylates. The polymeric material may be biodegradable and/or biocompatible. Non-limiting examples of specific polymers include poly(caprolactone) (PCL), ethylene vinyl acetate polymer (EVA), poly(lactic acid) (PLA), poly(L-lactic acid) (PLLA), poly(glycolic acid) (PGA), poly(lactic acid-co-glycolic acid) (PLGA), poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide) (PDLA), poly(L-lactide) (PLLA), poly(D,L-lactide-co-caprolactone), poly(D,L-lactide-co-caprolactone-co-glycolide), poly(D,L-lactide-co-PEO-co-D,L-lactide), poly(D,L-lactide-co-PPO-co-D,L-lactide), polyalkyl cyanoacralate, polyurethane, poly-L-lysine (PLL), hydroxypropyl methacrylate (HPMA), polyethyleneglycol, poly-L-glutamic acid, poly(hydroxy acids), polyanhydrides, polyorthoesters, poly(ester amides), polyamides, poly(ester ethers), polycarbonates, polyalkylenes such as polyethylene and polypropylene, polyalkylene glycols such as poly(ethylene glycol) (PEG), polyalkylene oxides (PEO), polyalkylene terephthalates such as poly(ethylene terephthalate), polyvinyl alcohols (PVA), polyvinyl ethers, polyvinyl esters such as poly(vinyl acetate), polyvinyl halides such as poly(vinyl chloride) (PVC), polyvinylpyrrolidone, polysiloxanes, polystyrene (PS), polyurethanes, derivatized celluloses such as alkyl celluloses, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitro celluloses, hydroxypropylcellulose, carboxymethylcellulose, polymers of acrylic acids, such as poly(methyl(meth)acrylate) (PMMA), poly(ethyl(meth)acrylate), poly(butyl(meth)acrylate), poly(isobutyl(meth)acrylate), poly(hexyl(meth)acrylate), poly(isodecyl(meth)acrylate), poly(lauryl(meth)acrylate), poly(phenyl(meth)acrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) and copolymers and mixtures thereof, polydioxanone and its copolymers, polyhydroxyalkanoates, polypropylene fumarate, polyoxymethylene, poloxamers, poly(ortho)esters, poly(butyric acid), poly(valeric acid), poly(lactide-co-caprolactone), and trimethylene carbonate, polyvinylpyrrolidone. The lipid nanoparticle may be coated or associated with a co-polymer such as, but not limited to, a block co-polymer, and (poly(ethylene glycol))-(poly(propylene oxide))-(poly(ethylene glycol))triblock copolymer (see US Publication 20120121718 and US Publication 20100003337; herein incorporated by reference in their entireties).

The vitamin of the polymer-vitamin conjugate may be vitamin E. The vitamin portion of the conjugate may be substituted with other suitable components such as, but not limited to, vitamin A, vitamin E, other vitamins, cholesterol, a hydrophobic moiety, or a hydrophobic component of other surfactants (e.g., sterol chains, fatty acids, hydrocarbon chains and alkylene oxide chains).

The lipid nanoparticle engineered to penetrate mucus may include surface altering agents such as, but not limited to, mmRNA, anionic protein (e.g., bovine serum albumin), surfactants (e.g., cationic surfactants such as for example dimethyldioctadecyl-ammonium bromide), sugars or sugar derivatives (e.g., cyclodextrin), nucleic acids, polymers (e.g., heparin, polyethylene glycol and poloxamer), mucolytic agents (e.g., N-acetylcysteine, mugwort, bromelain, papain, clerodendrum, acetylcysteine, bromhexine, carbocisteine, eprazinone, mesna, ambroxol, sobrerol, domiodol, letosteine, stepronin, tiopronin, gelsolin, thymosin (34 dornase alfa, neltenexine, erdosteine) and various DNases including rhDNase. The surface altering agent may be embedded or enmeshed in the particle's surface or disposed (e.g., by coating, adsorption, covalent linkage, or other process) on the surface of the lipid nanoparticle. (see US Publication 20100215580 and US Publication 20080166414; herein incorporated by reference in their entireties).

The mucus penetrating lipid nanoparticles may comprise at least one signal-sensor polynucleotide, primary construct, or mmRNA described herein. The signal-sensor polynucleotide, primary construct, or mmRNA may be encapsulated in the lipid nanoparticle and/or disposed on the surface of the particle. The signal-sensor polynucleotide, primary construct, or mmRNA may be covalently coupled to the lipid nanoparticle. Formulations of mucus penetrating lipid nanoparticles may comprise a plurality of nanoparticles. Further, the formulations may contain particles which may interact with the mucus and alter the structural and/or adhesive properties of the surrounding mucus to decrease mucoadhesion which may increase the delivery of the mucus penetrating lipid nanoparticles to the mucosal tissue.

In one embodiment, the nanoparticle may be for a dual modality therapy such as described by Mieszawska et al. (Bioconjugate Chemistry, 2013, 24 (9), pp 1429-1434; the contents of which is herein incorporated by reference in its entirety) comprising at least one therapeutic agent (e.g., a signal-sequence polynucleotide described herein). The therapeutic agent or agents formulated in the lipid nanoparticle may be an anti-angiogenic and a cytotoxic agent (see e.g., the polymer-lipid nanoparticles taught by Mieszawska et al. Bioconjugate Chemistry, 2013, 24 (9), pp 1429-1434; the contents of which is herein incorporated by reference in its entirety).

In another embodiment, the nanoparticle may comprise a LyP-1 peptide such as the nanocarrier composition described in International Patent Publication No. WO2013100869, the contents of which are herein incorporated by reference in its entirety. The LyP-1 peptide may be contained in the nanoparticles disclosed herein, or may be a conjugate, derivative, analogue or pegylated form of the peptide. In one embodiment, a nanoparticle comprising the LyP-1 peptide may comprise a signal-sensor polynucleotide and may be used for cancer treatment and/or imaging.

In one embodiment, the signal-sensor polynucleotide, primary construct, or mmRNA is formulated as a lipoplex, such as, without limitation, the ATUPLEX™ system, the DACC system, the DBTC system and other siRNA-lipoplex technology from Silence Therapeutics (London, United Kingdom), STEMFECT™ from STEMGENT® (Cambridge, Mass.), and polyethylenimine (PEI) or protamine-based targeted and non-targeted delivery of nucleic acids acids (Aleku et al. Cancer Res. 2008 68:9788-9798; Strumberg et al. Int J Clin Pharmacol Ther 2012 50:76-78; Santel et al., Gene Ther 2006 13:1222-1234; Santel et al., Gene Ther 2006 13:1360-1370; Gutbier et al., Pulm Pharmacol. Ther. 2010 23:334-344; Kaufmann et al. Microvasc Res 2010 80:286-293 Weide et al. J Immunother. 2009 32:498-507; Weide et al. J Immunother. 2008 31:180-188; Pascolo Expert Opin. Biol. Ther. 4:1285-1294; Fotin-Mleczek et al., 2011 J. Immunother. 34:1-15; Song et al., Nature Biotechnol. 2005, 23:709-717; Peer et al., Proc Natl Acad Sci USA. 2007 6; 104:4095-4100; deFougerolles Hum Gene Ther. 2008 19:125-132; all of which are incorporated herein by reference in its entirety).

In one embodiment such formulations may also be constructed or compositions altered such that they passively or actively are directed to different cell types in vivo, including but not limited to hepatocytes, immune cells, tumor cells, endothelial cells, antigen presenting cells, and leukocytes (Akinc et al. Mol Ther. 2010 18:1357-1364; Song et al., Nat Biotechnol. 2005 23:709-717; Judge et al., J Clin Invest. 2009 119:661-673; Kaufmann et al., Microvasc Res 2010 80:286-293; Santel et al., Gene Ther 2006 13:1222-1234; Santel et al., Gene Ther 2006 13:1360-1370; Gutbier et al., Pulm Pharmacol. Ther. 2010 23:334-344; Basha et al., Mol. Ther. 2011 19:2186-2200; Fenske and Cullis, Expert Opin Drug Deliv. 2008 5:25-44; Peer et al., Science. 2008 319:627-630; Peer and Lieberman, Gene Ther. 2011 18:1127-1133; all of which are incorporated herein by reference in its entirety). One example of passive targeting of formulations to liver cells includes the DLin-DMA, DLin-KC2-DMA and MC3-based lipid nanoparticle formulations which have been shown to bind to apolipoprotein E and promote binding and uptake of these formulations into hepatocytes in vivo (Akinc et al. Mol Ther. 2010 18:1357-1364; herein incorporated by reference in its entirety). Formulations can also be selectively targeted through expression of different ligands on their surface as exemplified by, but not limited by, folate, transferrin, N-acetylgalactosamine (GalNAc), and antibody targeted approaches (Kolhatkar et al., Curr Drug Discov Technol. 2011 8:197-206; Musacchio and Torchilin, Front Biosci. 2011 16:1388-1412; Yu et al., Mol Membr Biol. 2010 27:286-298; Patil et al., Crit Rev Ther Drug Carrier Syst. 2008 25:1-61; Benoit et al., Biomacromolecules. 2011 12:2708-2714 Zhao et al., Expert Opin Drug Deliv. 2008 5:309-319; Akinc et al., Mol Ther. 2010 18:1357-1364; Srinivasan et al., Methods Mol Biol. 2012 820:105-116; Ben-Arie et al., Methods Mol Biol. 2012 757:497-507; Peer 2010 J Control Release. 20:63-68; Peer et al., Proc Natl Acad Sci USA. 2007 104:4095-4100; Kim et al., Methods Mol Biol. 2011 721:339-353; Subramanya et al., Mol Ther. 2010 18:2028-2037; Song et al., Nat Biotechnol. 2005 23:709-717; Peer et al., Science. 2008 319:627-630; Peer and Lieberman, Gene Ther. 2011 18:1127-1133; all of which are incorporated herein by reference in its entirety).

In one embodiment, the signal-sensor polynucleotide, primary construct, or mmRNA is formulated as a solid lipid nanoparticle. A solid lipid nanoparticle (SLN) may be spherical with an average diameter between 10 to 1000 nm. SLN possess a solid lipid core matrix that can solubilize lipophilic molecules and may be stabilized with surfactants and/or emulsifiers. In a further embodiment, the lipid nanoparticle may be a self-assembly lipid-polymer nanoparticle (see Zhang et al., ACS Nano, 2008, 2 (8), pp 1696-1702; herein incorporated by reference in its entirety).

Liposomes, lipoplexes, or lipid nanoparticles may be used to improve the efficacy of signal-sensor polynucleotide, primary construct, or mmRNA directed protein production as these formulations may be able to increase cell transfection by the signal-sensor polynucleotide, primary construct, or mmRNA; and/or increase the translation of encoded protein. One such example involves the use of lipid encapsulation to enable the effective systemic delivery of polyplex plasmid DNA (Heyes et al., Mol Ther. 2007 15:713-720; herein incorporated by reference in its entirety). The liposomes, lipoplexes, or lipid nanoparticles may also be used to increase the stability of the signal-sensor polynucleotide, primary construct, or mmRNA.

Polymers, Biodegradable Nanoparticles, and Core-Shell Nanoparticles

The signal-sensor polynucleotide, primary construct, and mmRNA of the invention can be formulated using natural and/or synthetic polymers. Non-limiting examples of polymers which may be used for delivery include, but are not limited to, Dynamic POLYCONJUGATE™ formulations from MIRUS® Bio (Madison, Wis.) and Roche Madison (Madison, Wis.), PHASERX™ polymer formulations such as, without limitation, SMARTT POLYMER TECHNOLOGY™ (Seattle, Wash.), DMRI/DOPE, poloxamer, VAXFECTIN® adjuvant from Vical (San Diego, Calif.), chitosan, cyclodextrin from Calando Pharmaceuticals (Pasadena, Calif.), dendrimers and poly(lactic-co-glycolic acid) (PLGA) polymers. RONDEL™ (RNAi/Oligonucleotide Nanoparticle Delivery) polymers (Arrowhead Research Corporation, Pasadena, Calif.) and pH responsive co-block polymers such as, but not limited to, PHASERX™ (Seattle, Wash.).

A non-limiting example of PLGA formulations include, but are not limited to, PLGA injectable depots (e.g., ELIGARD® which is formed by dissolving PLGA in 66% N-methyl-2-pyrrolidone (NMP) and the remainder being aqueous solvent and leuprolide. Once injected, the PLGA and leuprolide peptide precipitates into the subcutaneous space).

Many of these polymer approaches have demonstrated efficacy in delivering oligonucleotides in vivo into the cell cytoplasm (reviewed in deFougerolles Hum Gene Ther. 2008 19:125-132; herein incorporated by reference in its entirety). Two polymer approaches that have yielded robust in vivo delivery of nucleic acids, in this case with small interfering RNA (siRNA), are dynamic polyconjugates and cyclodextrin-based nanoparticles. The first of these delivery approaches uses dynamic polyconjugates and has been shown in vivo in mice to effectively deliver siRNA and silence endogenous target mRNA in hepatocytes (Rozema et al., Proc Natl Acad Sci USA. 2007 104:12982-12887). This particular approach is a multicomponent polymer system whose key features include a membrane-active polymer to which nucleic acid, in this case siRNA, is covalently coupled via a disulfide bond and where both PEG (for charge masking) and N-acetylgalactosamine (for hepatocyte targeting) groups are linked via pH-sensitive bonds (Rozema et al., Proc Natl Acad Sci USA. 2007 104:12982-12887). On binding to the hepatocyte and entry into the endosome, the polymer complex disassembles in the low-pH environment, with the polymer exposing its positive charge, leading to endosomal escape and cytoplasmic release of the siRNA from the polymer. Through replacement of the N-acetylgalactosamine group with a mannose group, it was shown one could alter targeting from asialoglycoprotein receptor-expressing hepatocytes to sinusoidal endotheliu