Targeted exon skipping to correct exon duplications in the dystrophin gene

K.L. Greer; H. Lochmüller; K. Flanigan; S. Fletcher; S.D. Wilton

doi:10.1038/mtna.2014.8

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Targeted exon skipping to correct exon duplications in the dystrophin gene

Journal article

Open access

Peer reviewed

Targeted exon skipping to correct exon duplications in the dystrophin gene

K.L. Greer, H. Lochmüller, K. Flanigan, S. Fletcher and S.D. Wilton

Molecular Therapy — Nucleic Acids, Vol.3(3), e155

2014

DOI: https://doi.org/10.1038/mtna.2014.8

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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.

Details

Title: Targeted exon skipping to correct exon duplications in the dystrophin gene
Authors/Creators: K.L. Greer (Author/Creator) - The University of Western Australia
H. Lochmüller (Author/Creator) - Centre for Life
K. Flanigan (Author/Creator) - Nationwide Children's Hospital
S. Fletcher (Author/Creator) - Murdoch University
S.D. Wilton (Author/Creator) - Murdoch University
Publication Details: Molecular Therapy — Nucleic Acids, Vol.3(3), e155
Publisher: Nature Publishing Group
Identifiers: 991005544058207891
Copyright: © 2014 The American Society of Gene & Cell Therapy
Murdoch Affiliation: Centre for Comparative Genomics
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.255 Musculoskeletal Disorders; 1.255.628 Duchenne Muscular Dystrophy
Web Of Science research areas: Medicine, Research & Experimental
ESI research areas: Biology & Biochemistry