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

Abstract

Duchenne muscular dystrophy (DMD), caused by mutations in the X-linked dystrophin gene (<i>DMD</i>), 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 <i>DMD</i> mutations in patient-derived induced pluripotent stem cells (iPSCs) and <i>mdx</i> mice, an animal model of DMD. Cpf1-mediated genomic editing of human iPSCs, either by skipping of an out-of-frame <i>DMD</i> 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 <i>mdx</i> 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.