title: In vivo gene editing in dystrophic mouse muscle and muscle stem cells.
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.
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA.|Biological and Biomedical Sciences Program, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Mohammadsharif
authors.initials: M
authors.lastname: Tabebordbar
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA.|Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
authors.forename: Kexian
authors.initials: K
authors.lastname: Zhu
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Jason K W
authors.initials: JKW
authors.lastname: Cheng
authors.affiliation: Biological and Biomedical Sciences Program, Harvard Medical School, Boston, MA 02115, USA.|Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Wei Leong
authors.initials: WL
authors.lastname: Chew
authors.affiliation: Division of Genetics and Program in Genomics, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Widrick
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.|McGovern Institute for Brain Research, Department of Brain and Cognitive Science, and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Winston X
authors.initials: WX
authors.lastname: Yan
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Claire
authors.initials: C
authors.lastname: Maesner
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Elizabeth Y
authors.initials: EY
authors.lastname: Wu
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA.
authors.forename: Ru
authors.initials: R
authors.lastname: Xiao
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.|McGovern Institute for Brain Research, Department of Brain and Cognitive Science, and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: F Ann
authors.initials: FA
authors.lastname: Ran
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.|McGovern Institute for Brain Research, Department of Brain and Cognitive Science, and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Le
authors.initials: L
authors.lastname: Cong
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.|McGovern Institute for Brain Research, Department of Brain and Cognitive Science, and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, 20 Staniford Street, Boston, MA 02114, USA.
authors.forename: Luk H
authors.initials: LH
authors.lastname: Vandenberghe
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: George M
authors.initials: GM
authors.lastname: Church
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Amy J
authors.initials: AJ
authors.lastname: Wagers
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D000229:Dependovirus; D004195:Disease Models, Animal; D005091:Exons; D016368:Frameshift Mutation; D015316:Genetic Therapy / Q000379:methods*; D051379:Mice; D018101:Mice, Inbred mdx; D018482:Muscle, Skeletal / Q000378:metabolism; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000628:therapy*; D009206:Myocardium / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D032496:Satellite Cells, Skeletal Muscle / Q000378:metabolism*; D017384:Sequence Deletion; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: D012333:RNA, Messenger