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 at least one oligomer to a patient, wherein the at least one oligomer comprises a sequence of:
(SEQ ID NO: 10)
a) CXG XXG CCX CCG GXX CXG AAG GXG XXC XXG;
or
(SEQ ID NO: 12)
b) XXG CCX CCG GXX CXG AAG GXG XXC XXG XAC;
wherein X=U or T, wherein the sequence of the at least one oligomer can vary from the above sequence at up to two base positions.
2. The method according to claim 1 , wherein at least two oligomers are administered to the patient and each of the at least two oligomers causes skipping of a different exon.
3. The method according to claim 1 , wherein at least two oligomers are administered to the patient and each of the at least two oligomers causes skipping of the same exon.
4. The method according to claim 1 , wherein the at least one oligomer includes at least one modified nucleotide.
5. The method according to claim 4 , wherein the at least one modified nucleotide is modified at the 2′ position of the ribose.
6. The method according to claim 5 , wherein the at least one modified nucleotide is a 2′-O-methyl oligonucleotide.
7. The method according to claim 4 , wherein the at least one modified nucleotide is a phosphorodiamidate morpholino oligonucleotide.
8. The method according to claim 1 , wherein the at least one oligomer is conjugated to or complexed with a targeting protein, wherein the targeting protein targets the at least one oligomer to muscle tissue.
9. The method according to claim 1 , wherein the at least one oligomer is conjugated to or complexed with an arginine-rich cell penetrating peptide.
10. The method according to claim 1 , comprising administering the at least one oligomer to a patient by injection.
11. The method according to claim 10 , wherein the injection is an intramuscular, intravenous or subcutaneous injection.
12. The method according to claim 1 , comprising administering the at least one oligomer to a patient by oral administration.
13. The method according to claim 12 , wherein the oral administration includes administration using a capsule, a tablet, an aqueous suspension and/or an aqueous solution.
14. The method according to claim 1 , wherein administration of the at least one 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 at least one oligomer.