METHODS AND MEANS FOR EFFICIENT SKIPPING OF EXON 45 IN DUCHENNE MUSCULAR DYSTROPHY PRE-mRNA
The invention relates to a method for inducing or promoting skipping of exon 45 of DMD pre-mRNA in a Duchenne Muscular Dystrophy patient, preferably in an isolated (muscle) cell, the method comprising providing an isolate muscle cell with a molecule that binds to a continuous stretch of at least 21 nucleotides within said exon. The invention further relates to such molecule used in the method.
1 . An isolated antisense oligonucleotide consisting of 22, 23, 24, 25, 26, 27, 28 or 29 nucleotides, wherein said oligonucleotide is complementary along its entire length to a sequence in part of the human dystrophin exon 45 pre-mRNA, wherein said sequence is complementary to at least 22 nucleotides of a sequence consisting of 5′-UUUGCCGCUGCCCAAUGCCAUCCUG-3′ (SEQ ID NO: 3).
2 . The oligonucleotide of claim 1 , wherein said oligonucleotide comprises a phosphorothioate internucleoside linkage and a 2′-O-alkyl substituted ribose moiety.
3 . The oligonucleotide of claim 1 , wherein said oligonucleotide induces skipping of exon 45.
4 . The oligonucleotide of claim 1 , wherein the oligonucleotide comprises a modified base, and/or a modified sugar moiety, and/or a modified internucleoside linkage.
5 . The oligonucleotide of claim 4 , wherein the modified sugar moiety is a ribose that is mono- or di-substituted at the 2′, 3′, and/or 5′ position.
6 . The oligonucleotide of claim 1 , wherein the oligonucleotide comprises a phosphorodiamidate morpholino oligomer (PMO), peptide nucleic acid, and/or locked nucleic acid.
7 . The oligonucleotide of claim 1 , wherein the nucleotides of said oligonucleotide comprise purine and pyrimidine bases.
8 . The oligonucleotide of claim 7 , wherein the bases are selected from the group consisting of: adenine, cytosine, guanine, thymine and uracil.