Method of inhibiting miR-33 using a modified oligonucleotide
Compounds, compositions and methods are provided for modulating the expression and function of small non-coding RNAs. The compositions comprise oligomeric compounds, targeted to small non-coding RNAs. Methods of using these compounds for modulation of small non-coding RNAs as well as downstream targets of these RNAs and for diagnosis and treatment of disease associated with small non-coding RNAs are also provided.
1. A method of inhibiting the activity, function, or amount of miR-33, comprising contacting a cell with a compound comprising a modified oligonucleotide, wherein:
the modified is 100% complementary to miR-33;
the modified oligonucleotide consists of 8, 9, 10, 11, or 12 linked monomeric subunits; and wherein each monomeric subunit comprises a modified sugar moiety.
2. The method of claim 1 , wherein the contacting comprises administering the compound to a subject.
3. The method of claim 1 , wherein the miR-33 has the sequence of SEQ ID NO: 227.
4. The method of claim 1 , wherein each modified sugar moiety is independently selected from a 2′-F sugar moiety, a 2′-O-methyl sugar moiety, a 2′-O-methoxyethyl sugar moiety, and a bicyclic sugar moiety.
5. The method of claim 4 , wherein the bicyclic sugar moiety has a 4′-CH 2 —O-2′ bridge.
6. The method of claim 1 , wherein the modified oligonucleotide comprises at least one modified internucleoside linkage.
7. The method of claim 6 , wherein the modified internucleoside linkage is a phosphorothioate linkage.
8. The method of claim 7 , wherein each internucleoside linkage of the modified is a phosphorothioate linkage.
9. The method of claim 1 , wherein the oligonucleotide comprises two or more chemically distinct regions.
10. The method of claim 9 , wherein the sugar moiety of each nucleoside of a first chemically distinct region is the same and is selected from a 2′-fluoro nucleoside, a 2′-O-methyl nucleoside, a 2′-O-methoxyethyl nucleoside, and a bicyclic sugar nucleoside.
11. The method of claim 10 , wherein the bicyclic nucleoside has a 4′-CH 2 —O-2′ bridge.
12. The method of claim 1 , wherein the oligonucleotide comprises at least one 5-methylcytosine.
13. The method of claim 1 wherein the modified oligonucleotide consists of 8 monomeric subunits.
14. The method of claim 1 wherein the modified oligonucleotide consists of 9 monomeric subunits.
15. The method of claim 1 wherein the modified oligonucleotide consists of 10 monomeric subunits.
16. The method of claim 1 wherein the modified oligonucleotide consists of 11 monomeric subunits.
17. The method of claim 1 wherein the modified oligonucleotide consists of 12 monomeric subunits.
18. The method of claim 1 , wherein the modified oligonucleotide is attached to a conjugate group.
19. The method of claim 18 , wherein the conjugate group is cholesterol.
20. The method of claim 18 , wherein the conjugate group comprises a carbohydrate.
21. The method of claim 1 , wherein each modified sugar moiety is a bicyclic sugar moiety.
22. The method of claim 1 , wherein each modified sugar moiety is a bicyclic sugar moiety having a 4′-CH 2 —O-2′ bridge.
23. The method of claim 1 , wherein the modified oligonucleotide is complementary to a target region comprising nucleobases 1 to 8 of miR-33.
24. The method of claim 1 , wherein the modified oligonucleotide is complementary to a target region comprising nucleobases 2 to 9 of miR-33.
25. The method of claim 1 , wherein the modified oligonucleotide is complementary to a target region comprising nucleobases 3 to 10 of miR-33.