Oligomers
Molecules are provided for inducing or facilitating exon skipping in forming spliced mRNA products from pre-mRNA molecules in cells. The molecules may be provided directly as oligonucleotides or expression products of vectors that are administered to a subject. High rates of skipping can be achieved. High rates of skipping reduce the severity of a disease like Duchene Muscular Dystrophy so that the disease is more like Becker Muscular Dystrophy. This is a severe reduction in symptom severity and mortality.
1. A method of ameliorating Duchenne muscular dystrophy, the method comprising administering an oligomer to a patient, wherein the oligomer comprises at least 25 nucleotides and has a nucleotide sequence that is:
a) identical to at least 25 contiguous nucleotides of CXG XXG CCX CCG GXX CXG AAG GXG XXC XXG (SEQ ID NO: 10); or
b) identical to at least 25 contiguous nucleotides of XXG CCX CCG GXX CXG AAG GXG XXC XXG XAC (SEQ ID NO: 12),
wherein X=U or T.
2. The method according to claim 1 , wherein the oligomer comprises at least 25 nucleotides and has a nucleotide sequence that is identical to at least 25 contiguous nucleotides of CXG XXG CCX CCG GXX CXG AAG GXG XXC XXG (SEQ ID NO: 10).
3. The method according to claim 1 , wherein the oligomer comprises at least 25 nucleotides and has a nucleotide sequence that is identical to at least 25 contiguous nucleotides of XXG CCX CCG GXX CXG AAG GXG XXC XXG XAC (SEQ ID NO: 12).
4. The method according to claim 1 , wherein the oligomer is between 25 and 30 nucleotides in length.
5. The method according to claim 1 , wherein the oligomer is 25 nucleotides in length.
6. The method according to claim 1 , wherein the oligomer is 30 nucleotides in length.
7. The method according to claim 1 , wherein the method includes administering at least two oligomers and each of the at least two oligomers causes skipping of a different exon.
8. The method according to claim 1 , wherein the method includes administering at least two oligomers and each of the at least two oligomers causes skipping of the same exon.
9. The method according to claim 1 , wherein the oligomer includes at least one modified nucleotide.
10. The method according to claim 9 , wherein the at least one modified nucleotide is modified at the 2′ position of the ribose.
11. The method according to claim 10 , wherein the at least one modified nucleotide is a 2′-O-methyl oligonucleotide.
12. The method according to claim 9 , wherein the at least one modified nucleotide is a phosphorodiamidate morpholino oligonucleotide.
13. The method according to claim 1 , wherein the oligomer is conjugated to or complexed with a targeting protein, wherein the targeting protein targets the at least one oligomer to muscle tissue.
14. The method according to claim 1 , wherein the oligomer is conjugated to or complexed with an arginine-rich cell penetrating peptide.
15. The method according to claim 1 , comprising administering the oligomer to a patient by injection.
16. The method according to claim 15 , wherein the injection is an intramuscular, intravenous or subcutaneous injection.
17. The method according to claim 1 , comprising administering the oligomer to a patient by oral administration.
18. The method according to claim 17 , wherein the oral administration includes administration using a capsule, a tablet, an aqueous suspension and/or an aqueous solution.
19. The method according to claim 1 , wherein administration of the oligomer results in increased expression of a truncated dystrophin protein in a muscle of the patient as compared to expression of the truncated dystrophin protein in the muscle of the patient prior to administration of the oligomer.