Micrornas that regulate muscle cell proliferation and differentiation
View Patent ↗The presently disclosed subject matter provides methods and compositions for modulating gene expression in myocytes. Also provided are cells comprising the compositions of the presently disclosed subject matter.
1. A method for treating a muscle injury in a subject comprising administering to a muscle injury site in the subject a combination of miR-133, wherein miR-133 comprises the nucleotide sequence of SEQ ID NO: 2, and an inhibitor of miR-1, wherein the inhibitor is a polynucleotide comprising a nucleotide sequence that is complementary to SEQ ID NO: 1.
2. The method of claim 1 , further comprising administering to the muscle injury site at a subsequent time point a combination of miR-1, wherein miR-1 comprises the nucleotide sequence of SEQ ID NO: 1, and an inhibitor of miR-133, wherein the inhibitor is a polynucleotide comprising a nucleotide sequence that is complementary to SEQ ID NO: 2.
3. The method of claim 1 , wherein the muscle injury results from a mechanical muscle trauma, a muscular degenerative disorder, a cardiac insult, or a combination thereof.
4. The method of claim 1 , wherein the subject is a mammal.
5. The method of claim 1 , wherein the inhibitor of miR-1 is a modified polynucleotide.
6. The method of claim 5 , wherein the modified polynucleotide is a 2′-O-methyl polynucleotide.
7. The method of claim 6 , wherein the inhibitor of miR-1 is 2′-O-methyl-miR-1 (SEQ ID NO: 62).
8. The method of claim 2 , wherein the inhibitor of miR-133 is a modified polynucleotide.
9. The method of claim 8 , wherein the modified polynucleotide is a 2′-O-methyl polynucleotide.
10. The method of claim 9 , wherein the inhibitor of miR-133 is 2′-O-methyl-miR-133 (SEQ ID NO: 63).
11. The method of claim 1 , wherein miR-133 is encoded by a vector.
12. The method of claim 11 , wherein the vector comprises:
(a) a promoter operatively linked to a nucleic acid molecule encoding miR-133; and
(b) a transcription termination sequence.
13. A method for treating a muscle injury in a subject comprising administering to a muscle injury site in the subject a combination of miR-1, wherein miR-1 comprises the nucleotide sequence of SEQ ID NO: 1, and an inhibitor of miR-133, wherein the inhibitor is a polynucleotide comprising a nucleotide sequence that is complementary to SEQ ID NO: 2.
14. The method of claim 13 , wherein the subject is a mammal.
15. An in vitro method for inducing myocyte differentiation comprising contacting a myocyte with miR-1, wherein miR-1 comprises the nucleotide sequence of SEQ ID NO: 1, and an inhibitor of miR-133, wherein the inhibitor is a polynucleotide comprising a nucleotide sequence that is complementary to SEQ ID NO: 2, wherein differentiation of the myocyte is induced.
16. The method of claim 15 , wherein miR-1 is encoded by a vector.
17. An in vitro method for promoting myocyte proliferation comprising contacting a myocyte with miR-133, wherein miR-133 comprises the nucleotide sequence of SEQ ID NO: 2 and an inhibitor of miR-1, wherein the inhibitor is a polynucleotide comprising nucleotide sequence that is complementary to SEQ ID NO: 1, wherein proliferation of the myocyte is promoted.
18. The method of claim 17 , wherein miR-133 is encoded by a vector.
19. The method of claim 2 , wherein miR-1 is encoded by a vector.
20. The method of claim 19 , wherein the vector comprises:
(a) a promoter operatively linked to a nucleic acid molecule encoding miR-1; and
(b) a transcription termination sequence.