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Targeted exon skipping to correct exon duplications in the dystrophin gene

Greer, K.L., Lochmüller, H., Flanigan, K., Fletcher, S. and Wilton, S.D. (2014) Targeted exon skipping to correct exon duplications in the dystrophin gene. Molecular Therapy — Nucleic Acids, 3 (3). e155.

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Abstract

Duchenne muscular dystrophy is a severe muscle-wasting disease caused by mutations in the dystrophin gene that ablate functional protein expression. Although exonic deletions are the most common Duchenne muscular dystrophy lesion, duplications account for 10–15% of reported disease-causing mutations, and exon 2 is the most commonly duplicated exon. Here, we describe the in vitro evaluation of phosphorodiamidate morpholino oligomers coupled to a cell-penetrating peptide and 2′-O-methyl phosphorothioate oligonucleotides, using three distinct strategies to reframe the dystrophin transcript in patient cells carrying an exon 2 duplication. Differences in exon-skipping efficiencies in vitro were observed between oligomer analogues of the same sequence, with the phosphorodiamidate morpholino oligomer coupled to a cell-penetrating peptide proving the most effective. Differences in exon 2 excision efficiency between normal and exon 2 duplication cells, were apparent, indicating that exon context influences oligomer-induced splice switching. Skipping of a single copy of exon 2 was induced in the cells carrying an exon 2 duplication, the simplest strategy to restore the reading frame and generate a normal dystrophin transcript. In contrast, multiexon skipping of exons 2–7 to generate a Becker muscular dystrophy-like dystrophin transcript was more challenging and could only be induced efficiently with the phosphorodiamidate morpholino oligomer chemistry.

Publication Type: Journal Article
Murdoch Affiliation: Centre for Comparative Genomics
Publisher: Nature Publishing Group
Copyright: © 2014 The American Society of Gene & Cell Therapy
URI: http://researchrepository.murdoch.edu.au/id/eprint/22096
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