Method for efficient exon (44) skipping in duchenne muscular dystrophy and associated means

Abstract

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 20 to 25 nucleotides in length that comprises a base sequence that binds and/or is complementary to SEQ ID NO: 3, 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: 39 or SEQ ID NO: 40. 
     
     
         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).