Method for efficient exon (44) skipping in duchenne muscular dystrophy and associated means
The invention relates to a nucleic acid molecule that binds and/or is complementary to the nucleotide molecule having sequence 5′-GUGGCUAACAGAAGCU (SEQ ID NO 1) and to its use in a method for inducing skipping of exon 44 of the DMD gene in a DMD patient.
1. A single stranded antisense oligonucleotide 16 to 25 nucleotides in length that comprises a base sequence of the sequence selected from the group consisting of SEQ ID NO: 5 to SEQ ID NO: 16, SEQ ID NO: 20 to SEQ ID NO: 28, SEQ ID NO: 30 to SEQ ID NO: 34, SEQ ID NO: 41, and SEQ ID NO: 46, wherein the antisense oligonucleotide induces skipping of exon 44 of human dystrophin pre-mRNA, and comprises a modification.
2. The antisense oligonucleotide according to claim 1 , wherein said antisense oligonucleotide comprises the base sequence of the sequence SEQ ID NO: 5.
3. The antisense oligonucleotide according to claim 1 , comprising a 2′-O-alkyl phosphorothioate antisense oligonucleotide.
4. The antisense oligonucleotide according to claim 3 , wherein said antisense oligonucleotide comprises a 2′-O-methyl phosphorothioate ribose.
5. A viral-based vector, comprising a Pol III-promoter driven expression cassette for expression of an antisense oligonucleotide according to claim 1 .
6. The antisense oligonucleotide of claim 1 , wherein said modification comprises a modified backbone.
7. The antisense oligonucleotide according to claim 6 , wherein the modified backbone is selected from the group consisting of a morpholino backbone, a carbamate backbone, a siloxane backbone, a sulfide backbone, a sulfoxide backbone, a sulfone backbone, a formacetyl backbone, a thioformacetyl backbone, a methyleneformacetyl backbone, a riboacetyl backbone, an alkene containing backbone, a sulfamate backbone, a sulfonate backbone, a sulfonamide backbone, a methyleneimino backbone, a methylenehydrazino backbone and an amide backbone.
8. The antisense oligonucleotide according to claim 1 , wherein said modification is selected from the group consisting of: phosphorodiamidate morpholino oligomer (PMO), peptide nucleic acid, and locked nucleic acid.
9. The antisense oligonucleotide according to claim 2 , wherein the antisense oligonucleotide is a PMO.
10. The oligonucleotide of claim 1 , wherein said oligonucleotide is conjugated to a ligand.
11. The oligonucleotide of claim 10 , wherein said ligand is a peptide.
12. The antisense oligonucleotide according to claim 1 , wherein said modification comprises a locked nucleic acid (LNA).