• Benjamin L Duchêne, Khadija Cherif, Jean-Paul Iyombe-Engembe, Antoine Guyon, Joel Rousseau, Dominique L Ouellet, Xavier Barbeau, Patrick Lague, and Jacques P Tremblay.
• Centre de Recherche du Centre Hospitalier Universitaire de Québec, Neurosciences Axis, Québec City, QC, Canada; Faculty of Medicine, Department of Molecular Medicine, Université Laval, Quebec City, QC, Canada.
• Mol. Ther. 2018 Nov 7; 26 (11): 2604-2616.

AbstractDuchenne muscular dystrophy (DMD), a severe hereditary disease affecting 1 in 3,500 boys, mainly results from the deletion of exon(s), leading to a reading frameshift of the DMD gene that abrogates dystrophin protein synthesis. Pairs of sgRNAs for the Cas9 of Staphylococcus aureus were meticulously chosen to restore a normal reading frame and also produce a dystrophin protein with normally phased spectrin-like repeats (SLRs), which is not usually obtained by skipping or by deletion of complete exons. This can, however, be obtained in rare instances where the exon and intron borders of the beginning and the end of the complete deletion (patient deletion plus CRISPR-induced deletion) are at similar positions in the SLR. We used pairs of sgRNAs targeting exons 47 and 58, and a normal reading frame was restored in myoblasts derived from muscle biopsies of 4 DMD patients with different exon deletions. Restoration of the DMD reading frame and restoration of dystrophin expression were also obtained in vivo in the heart of the del52hDMD/mdx. Our results provide a proof of principle that SaCas9 could be used to edit the human DMD gene and could be considered for further development of a therapy for DMD.Copyright © 2018 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.

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