In vivo gene editing in dystrophic mouse muscle and muscle stem cells.

Science
Authors
Keywords
Abstract

Frame-disrupting mutations in the DMD gene, encoding dystrophin, compromise myofiber integrity and drive muscle deterioration in Duchenne muscular dystrophy (DMD). Removing one or more exons from the mutated transcript can produce an in-frame mRNA and a truncated, but still functional, protein. In this study, we developed and tested a direct gene-editing approach to induce exon deletion and recover dystrophin expression in the mdx mouse model of DMD. Delivery by adeno-associated virus (AAV) of clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 endonucleases coupled with paired guide RNAs flanking the mutated Dmd exon23 resulted in excision of intervening DNA and restored the Dmd reading frame in myofibers, cardiomyocytes, and muscle stem cells after local or systemic delivery. AAV-Dmd CRISPR treatment partially recovered muscle functional deficiencies and generated a pool of endogenously corrected myogenic precursors in mdx mouse muscle.

Year of Publication
2016
Journal
Science
Volume
351
Issue
6271
Pages
407-11
Date Published
2016 Jan 22
ISSN
1095-9203
URL
DOI
10.1126/science.aad5177
PubMed ID
26721686
PubMed Central ID
PMC4924477
Links
Grant list
P50 HG005550 / HG / NHGRI NIH HHS / United States
R01 MH110049 / MH / NIMH NIH HHS / United States
5U01HL100402 / HL / NHLBI NIH HHS / United States
5DP1-MH100706 / DP / NCCDPHP CDC HHS / United States
T32 GM007598 / GM / NIGMS NIH HHS / United States
T32 GM007753 / GM / NIGMS NIH HHS / United States
T2GM007753 / GM / NIGMS NIH HHS / United States
U01 HL100402 / HL / NHLBI NIH HHS / United States
DP2 OD004345 / OD / NIH HHS / United States
DP1 MH100706 / MH / NIMH NIH HHS / United States
PN2 EY018244 / EY / NEI NIH HHS / United States
5PN2EY018244 / EY / NEI NIH HHS / United States
Howard Hughes Medical Institute / United States
R01 DK097768 / DK / NIDDK NIH HHS / United States
T32 GM008313 / GM / NIGMS NIH HHS / United States
1DP2OD004345 / OD / NIH HHS / United States
5R01DK097768-03 / DK / NIDDK NIH HHS / United States