Antisense oligonucleotides for inducing exon skipping and methods of use thereof
An antisense molecule capable of binding to a selected target site to induce exon skipping in the dystrophin gene, as set forth in SEQ ID NO: 1 to 202.
1. An isolated antisense oligonucleotide of 25 to 50 nucleotides in length comprising SEQ ID NO: 180, wherein the uracil bases are optionally thymine bases.
2. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide comprises a non-natural backbone.
3. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide is chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligonucleotide.
4. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide does not activate RNase H.
5. The antisense oligonucleotide of claim 2 , wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties.
6. The antisense oligonucleotide of claim 5 , wherein the non-natural moieties are morpholinos.
7. The antisense oligonucleotide of claim 6 , wherein the uracil bases are thymine bases.
8. The antisense oligonucleotide of claim 1 , wherein the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages.
9. The antisense oligonucleotide of claim 8 , wherein the non-natural inter-nucleotide linkages are modified phosphates.
10. The antisense oligonucleotide of claim 9 , wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates.
11. The antisense oligonucleotide of claim 10 , wherein the modified phosphates are phosphoroamidates.
12. The antisense oligonucleotide of claim 10 , wherein the modified phosphates are phosphoromorpholidates.
13. The antisense oligonucleotide of claim 1 , wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties and the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages.
14. The antisense oligonucleotide of claim 13 , wherein the non-natural moieties are morpholinos and the non-natural internucleotide linkages are modified phosphates.
15. The antisense oligonucleotide of claim 14 , wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates.
16. The antisense oligonucleotide of claim 15 , wherein the modified phosphates are phosphoroamidates.
17. The antisense oligonucleotide of claim 15 , wherein the modified phosphates are phosphoromorpholidates.
18. The antisense oligonucleotide of claim 15 , wherein the uracil bases are thymine bases.
19. The antisense oligonucleotide of claim 18 , wherein the antisense oligonucleotide is chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligonucleotide.
20. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide is 25 nucleotides in length.
21. The antisense oligonucleotide of claim 20 , wherein the uracil bases are thymine bases.
22. A pharmaceutical composition, comprising the antisense oligonucleotide of claim 1 and a pharmaceutically acceptable carrier.
23. The pharmaceutical composition of claim 22 , wherein the pharmaceutically acceptable carrier is phosphate-buffered saline.
24. A pharmaceutically acceptable salt of the antisense oligonucleotide of claim 1 .