Sci Adv. 2017 Apr 12;3(4):e1602814. doi: 10.1126/sciadv.1602814. eCollection 2017.

CRISPR-Cpf1 correction of muscular dystrophy mutations in human cardiomyocytes and mice.

Zhang Y^1,2,3, Long C^1,2,3, Li H^1,2,3, McAnally JR^1,2,3, Baskin KK^1,2,3, Shelton JM⁴, Bassel-Duby R^1,2,3, Olson EN^1,2,3.

Abstract

Duchenne muscular dystrophy (DMD), caused by mutations in the X-linked dystrophin gene (DMD), is characterized by fatal degeneration of striated muscles. Dilated cardiomyopathy is one of the most common lethal features of the disease. We deployed Cpf1, a unique class 2 CRISPR (clustered regularly interspaced short palindromic repeats) effector, to correct DMD mutations in patient-derived induced pluripotent stem cells (iPSCs) and mdx mice, an animal model of DMD. Cpf1-mediated genomic editing of human iPSCs, either by skipping of an out-of-frame DMD exon or by correcting a nonsense mutation, restored dystrophin expression after differentiation to cardiomyocytes and enhanced contractile function. Similarly, pathophysiological hallmarks of muscular dystrophy were corrected in mdx mice following Cpf1-mediated germline editing. These findings are the first to show the efficiency of Cpf1-mediated correction of genetic mutations in human cells and an animal disease model and represent a significant step toward therapeutic translation of gene editing for correction of DMD.