Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Sci Adv. 2017 Apr 12;3(4):e1602814. doi: 10.1126/sciadv.1602814. eCollection 2017 Apr.
Duchenne muscular dystrophy (DMD), caused by mutations in the X-linked dystrophin gene (), 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 mutations in patient-derived induced pluripotent stem cells (iPSCs) and mice, an animal model of DMD. Cpf1-mediated genomic editing of human iPSCs, either by skipping of an out-of-frame 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 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.
杜氏肌营养不良症（DMD）是由 X 连锁的肌营养不良蛋白基因（）突变引起的，其特征是横纹肌进行性退化。扩张型心肌病是该病最常见的致死特征之一。我们利用 Cpf1，一种独特的 II 类 CRISPR（成簇规律间隔短回文重复序列）效应物，纠正了患者来源的诱导多能干细胞（iPSC）和 DMD 动物模型（）中的突变。Cpf1 介导的人类 iPSC 基因组编辑，无论是通过跳过框架外的外显子还是纠正无义突变，在分化为心肌细胞后均可恢复肌营养不良蛋白的表达，并增强收缩功能。同样，在 Cpf1 介导的种系编辑后，在 小鼠中纠正了肌肉疾病的病理生理特征。这些发现首次表明 Cpf1 介导的人类细胞和动物疾病模型中基因突变的校正效率，代表了基因编辑治疗 DMD 的校正向治疗转化的重要一步。
