CRISPR-Cas9 corrects Duchenne muscular dystrophy exon 44 deletion mutations in mice and human cells
CRISPR-Cas9 corrects Duchenne muscular dystrophy exon 44 deletion mutations in mice and human cells
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DOI：
10.1126/sciadv.aav4324
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发表时间：
2019-03-01
期刊：
影响因子：
13.6
通讯作者：
Olson, Eric N.
中科院分区：
文献类型：
--
作者：
Min, Yi-Li;Li, Hui;Olson, Eric N.
肌营养不良蛋白基因突变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变性。删除肌营养不良蛋白基因外显子44的突变是DMD最常见的原因之一，可以在大约12%的患者中通过编辑周围的外显子来纠正，从而恢复肌营养不良蛋白的开放阅读框。在这里，我们提出了一种简单有效的策略，通过CRISPR-Cas9基因编辑在患者来源的诱导多能干细胞和具有相同缺失突变的新小鼠模型中获得的心肌细胞中纠正外显子44缺失突变。利用AAV9编码Cas9和单导rna，我们还证明了这些基因编辑成分的剂量对体内最佳基因校正的重要性。我们的研究结果代表了基因编辑在临床应用中纠正DMD的重要一步。
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 similar to 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.