Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Sci Adv. 2021 Apr 30;7(18). doi: 10.1126/sciadv.abg4910. Print 2021 Apr.
Duchenne muscular dystrophy (DMD) is a fatal muscle disease caused by the lack of dystrophin, which maintains muscle membrane integrity. We used an adenine base editor (ABE) to modify splice donor sites of the dystrophin gene, causing skipping of a common DMD deletion mutation of exon 51 (∆Ex51) in cardiomyocytes derived from human induced pluripotent stem cells, restoring dystrophin expression. Prime editing was also capable of reframing the dystrophin open reading frame in these cardiomyocytes. Intramuscular injection of ∆Ex51 mice with adeno-associated virus serotype-9 encoding ABE components as a split-intein trans-splicing system allowed gene editing and disease correction in vivo. Our findings demonstrate the effectiveness of nucleotide editing for the correction of diverse DMD mutations with minimal modification of the genome, although improved delivery methods will be required before these strategies can be used to sufficiently edit the genome in patients with DMD.
杜氏肌营养不良症(DMD)是一种致命的肌肉疾病,由肌营养不良蛋白的缺失引起,肌营养不良蛋白维持肌肉细胞膜的完整性。我们使用腺嘌呤碱基编辑器(ABE)修饰肌营养不良蛋白基因的剪接供体部位,导致源自人诱导多能干细胞的心肌细胞中第 51 外显子(∆Ex51)的常见 DMD 缺失突变跳跃,恢复肌营养不良蛋白的表达。Prime 编辑也能够重新构建这些心肌细胞中的肌营养不良蛋白开放阅读框。腺相关病毒血清型 9 编码 ABE 组件作为分裂内含肽转剪接系统的肌内注射允许在体内进行基因编辑和疾病纠正。我们的研究结果表明,核苷酸编辑对于纠正多种 DMD 突变是有效的,同时对基因组的最小修饰,尽管在这些策略能够用于充分编辑 DMD 患者的基因组之前,需要改进的传递方法。