METHODS AND MEANS FOR EFFICIENT SKIPPING OF EXON 45 IN DUCHENNE MUSCULAR DYSTROPHY PRE-mRNA

Abstract

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.

We claim: 
     
         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.