Department of Molecular Biology, Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA.
Sen. Paul D. Wellstone Muscular Dystrophy Cooperative Research Center, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA.
Sci Adv. 2019 Mar 6;5(3):eaav4324. doi: 10.1126/sciadv.aav4324. eCollection 2019 Mar.
Mutations in the dystrophin gene cause Duchenne muscular dystrophy (DMD), which is characterized by lethal degeneration of cardiac and skeletal muscles. Mutations that delete exon 44 of the dystrophin gene represent one of the most common causes of DMD and can be corrected in ~12% of patients by editing surrounding exons, which restores the dystrophin open reading frame. Here, we present a simple and efficient strategy for correction of exon 44 deletion mutations by CRISPR-Cas9 gene editing in cardiomyocytes obtained from patient-derived induced pluripotent stem cells and in a new mouse model harboring the same deletion mutation. Using AAV9 encoding Cas9 and single guide RNAs, we also demonstrate the importance of the dosages of these gene editing components for optimal gene correction in vivo. Our findings represent a significant step toward possible clinical application of gene editing for correction of DMD.
肌营养不良蛋白基因突变会导致杜氏肌营养不良症（DMD），其特征是心脏和骨骼肌的致死性退化。肌营养不良蛋白基因外显子 44 缺失突变是 DMD 最常见的原因之一，通过编辑周围的外显子可以纠正约 12%的患者，从而恢复肌营养不良蛋白开放阅读框。在这里，我们提出了一种简单有效的策略，通过 CRISPR-Cas9 基因编辑，在从患者来源的诱导多能干细胞中获得的心肌细胞和携带相同缺失突变的新型小鼠模型中，纠正外显子 44 缺失突变。使用编码 Cas9 和单引导 RNA 的 AAV9，我们还证明了这些基因编辑成分的剂量对于体内最佳基因纠正的重要性。我们的研究结果为基因编辑纠正 DMD 的可能临床应用迈出了重要的一步。
