Статья

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

Mohammadsharif TabebordbarBiological and Biomedical Sciences Program, Harvard Medical School, Boston, MA 02115, USAKexian ZhuDepartment of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USAJason ChengDepartment of Stem Cell and Regenerative Biology, Harvard University, and Harvard Stem Cell Institute, Cambridge, MA 02138, USAWei Leong ChewBiological and Biomedical Sciences Program, Harvard Medical School, Boston, MA 02115, USAJeffrey J. WidrickDivision of Genetics and Program in Genomics, Boston Children’s Hospital, Harvard Medical School, Boston, MA 02115, USAWinston X. YanBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAClaire MaesnerDepartment of Stem Cell and Regenerative Biology, Harvard University, and Harvard Stem Cell Institute, Cambridge, MA 02138, USAElizabeth Y. WuDepartment of Stem Cell and Regenerative Biology, Harvard University, and Harvard Stem Cell Institute, Cambridge, MA 02138, USARu XiaoGrousbeck Gene Therapy Center, Schepens Eye Research Institute, and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USAF. Ann RanBroad Institute of MIT and Harvard, Cambridge, MA 02142, USALe CongBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAFeng ZhangBroad Institute of MIT and Harvard, Cambridge, MA 02142, USALuk H. VandenbergheGrousbeck Gene Therapy Center, Schepens Eye Research Institute, and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USAGeorge M. ChurchDepartment of Genetics, Harvard Medical School, Boston, MA 02115, USAAmy J. WagersDepartment of Stem Cell and Regenerative Biology, Harvard University, and Harvard Stem Cell Institute, Cambridge, MA 02138, USA

2015en

ABI

Аннотация

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.

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Идентификаторы

DOI: 10.1126/science.aad5177

Цитирования и источники

Цитирований: 2Использованных источников: 0

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