Single Exon Skipping Can Address a Multi-Exon Duplication in the Dystrophin Gene
Open Access
- 25 June 2020
- journal article
- research article
- Published by MDPI AG in International Journal of Molecular Sciences
- Vol. 21 (12), 4511
- https://doi.org/10.3390/ijms21124511
Abstract
Duchenne muscular dystrophy (DMD) is a severe muscle wasting disease typically caused by protein-truncating mutations that preclude synthesis of a functional dystrophin. Exonic deletions are the most common type of DMD lesion, however, whole exon duplications account for between 10–15% of all reported mutations. Here, we describe in vitro evaluation of antisense oligonucleotide-induced splice switching strategies to re-frame the transcript disrupted by a multi-exon duplication within the DMD gene. Phosphorodiamidate morpholino oligomers and phosphorodiamidate morpholino oligomers coupled to a cell penetrating peptide were evaluated in a Duchenne muscular dystrophy patient cell strain carrying an exon 14–17 duplication. Two strategies were employed; the conventional approach was to remove both copies of exon 17 in addition to exon 18, and the second strategy was to remove only the first copy of exon 17. Both approaches result in a larger than normal but in-frame DMD transcript, but surprisingly, the removal of only the first exon 17 appeared to be more efficient in restoring dystrophin, as determined using western blotting. The emergence of a normal sized DMD mRNA transcript that was not apparent in untreated samples may have arisen from back splicing and could also account for some of the dystrophin protein being produced.Funding Information
- National Institutes of Health (2R01 NS044146-05A1)
- National Health and Medical Research Council (1086311 and 1144791)
- Muscular Dystrophy Association (4352)
This publication has 36 references indexed in Scilit:
- Personalized exon skipping strategies to address clustered non-deletion dystrophin mutationsNeuromuscular Disorders, 2010
- By‐passing the nonsense mutation in the 4CV mouse model of muscular dystrophy by induced exon skippingThe Journal of Gene Medicine, 2008
- Antisense-induced exon skipping for duplications in Duchenne muscular dystrophyBMC Medical Genetics, 2007
- Antisense Oligonucleotide-induced Exon Skipping Across the Human Dystrophin Gene TranscriptMolecular Therapy, 2007
- Antisense oligonucleotide induced exon skipping and the dystrophin gene transcript: cocktails and chemistriesBMC Molecular Biology, 2007
- The Influence of Antisense Oligonucleotide Length on Dystrophin Exon SkippingMolecular Therapy, 2007
- Characterization of RNase R-digested cellular RNA source that consists of lariat and circular RNAs from pre-mRNA splicingNucleic Acids Research, 2006
- Functional Analysis of 114 Exon-Internal AONs for Targeted DMD Exon Skipping: Indication for Steric Hindrance of SR Protein Binding SitesOligonucleotides, 2005
- Improved antisense oligonucleotide induced exon skipping in the mdx mouse model of muscular dystrophyThe Journal of Gene Medicine, 2002
- Characterization of Dystrophin in Muscle-Biopsy Specimens from Patients with Duchenne's or Becker's Muscular DystrophyThe New England Journal of Medicine, 1988