Correction of the Exon 2 Duplication in DMD Myoblasts by a Single CRISPR/Cas9 System

• Published in: Molecular therapy. Nucleic acids Vol. 7; no. C; pp. 11 - 19
• Main Authors: Lattanzi, Annalisa, Duguez, Stephanie, Moiani, Arianna, Izmiryan, Araksya, Barbon, Elena, Martin, Samia, Mamchaoui, Kamel, Mouly, Vincent, Bernardi, Francesco, Mavilio, Fulvio, Bovolenta, Matteo
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16-06-2017; Elsevier Limited; Elsevier; American Society of Gene & Cell Therapy
• Subjects: Biochemistry, Molecular Biology; Clonal deletion; CRISPR; CRISPR/Cas9; Duchenne's muscular dystrophy; duplication; Dystrophin; Gene deletion; gene editing; Genetics; Genomes; gRNA; lentivirus; Life Sciences; Molecular biology; Muscular dystrophy; Mutation; Myoblasts; Myotubes; Original; Proteins; Reading; Ribonucleic acid; RNA; Transcription; France; dystrophin; CRISPR/Cas9; gene editing; duplication; lentivirus
• ISSN: 2162-2531; 2162-2531
• DOI: 10.1016/j.omtn.2017.02.004

Exonic duplications account for 10%–15% of all mutations in Duchenne muscular dystrophy (DMD), a severe hereditary neuromuscular disorder. We report a CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9-based strategy to correct the most frequent (exon 2) duplication in the DMD gene by targeted deletion, and tested the efficacy of such an approach in patient-derived myogenic cells. We demonstrate restoration of wild-type dystrophin expression at transcriptional and protein level in myotubes derived from genome-edited myoblasts in the absence of selection. Removal of the duplicated exon was achieved by the use of only one guide RNA (gRNA) directed against an intronic duplicated region, thereby increasing editing efficiency and reducing the risk of off-target effects. This study opens a novel therapeutic perspective for patients carrying disease-causing duplications.
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