Functional rescue of dystrophin deficiency in mice caused by frameshift mutations using Campylobacter jejuni Cas9

Koo, T; Lu-Nguyen, N B; Malerba, A; Kim, E; Kim, D; Cappellari, O; Cho, H-Y; Dickson, G; Popplewell, L; Kim, J-S

Abstract

Duchenne muscular dystrophy (DMD) is a fatal, X-linked muscle wasting disease caused by mutations in the DMD gene. In 51% of DMD cases, a reading frame is disrupted because of deletion of several exons. Here, we show that CjCas9 derived from Campylobacter jejuni can be
used as a gene editing tool to correct an out-of-frame Dmd exon in Dmd knockout mice. Herein, we used Cas9 derived from S. pyogenes to generate Dmd knockout (KO) mice with a frameshift mutation in Dmd gene. Then, we expressed CjCas9, its single-guide RNA, and the eGFP gene
in the tibialis anterior muscle of the Dmd KO mice using an all-in-one adeno-associated virus (AAV) vector. CjCas9 cleaved the target site in the Dmd gene efficiently in vivo and induced small insertions or deletions at the target site. This treatment resulted in conversion of the
disrupted Dmd reading frame from out-of-frame to in-frame, leading to the expression of dystrophin in the sarcolemma. Importantly, muscle strength was enhanced in the CjCas9-treated muscles, without off-target mutations, indicating high efficiency and specificity of CjCas9. This work suggests that in vivo DMD frame correction, mediated by CjCas9 has great potential for the treatment of DMD and other neuromuscular diseases.

Citation

Koo, T., Lu-Nguyen, N. B., Malerba, A., Kim, E., Kim, D., Cappellari, O., …Kim, J. (2018). Functional rescue of dystrophin deficiency in mice caused by frameshift mutations using Campylobacter jejuni Cas9. Molecular Therapy, https://doi.org/10.1016/j.ymthe.2018.03.018

• Journal Article Type: Article
• Acceptance Date: Mar 27, 2018
• Publication Date: Mar 30, 2018
• Deposit Date: Apr 11, 2018
• Publicly Available Date: May 30, 2018
• Journal: Molecular Therapy
• Print ISSN: 1525-0016
• Electronic ISSN: 1525-0024
• Publisher: Cell Press
• Peer Reviewed: Peer Reviewed
• DOI: https://doi.org/10.1016/j.ymthe.2018.03.018
• Public URL: https://rvc-repository.worktribe.com/output/1387901