清脆的
基因组编辑
骨骼肌
Cas9
生物
基因
心肌细胞
杜氏肌营养不良
强直性营养不良
遗传增强
肌营养不良
细胞生物学
遗传学
分子生物学
解剖
作者
Kiril Poukalov,M. Carmen Valero,Derek R. Muscato,Linnea M Adams,Heejae Chun,Youngil Lee,Nadja S. Andrade,Zane Zeier,H. Lee Sweeney,Eric T. Wang
标识
DOI:10.1073/pnas.2321438121
摘要
Successful CRISPR/Cas9-based gene editing in skeletal muscle is dependent on efficient propagation of Cas9 to all myonuclei in the myofiber. However, nuclear-targeted gene therapy cargos are strongly restricted to their myonuclear domain of origin. By screening nuclear localization signals and nuclear export signals, we identify “Myospreader,” a combination of short peptide sequences that promotes myonuclear propagation. Appending Myospreader to Cas9 enhances protein stability and myonuclear propagation in myoblasts and myofibers. AAV-delivered Myospreader dCas9 better inhibits transcription of toxic RNA in a myotonic dystrophy mouse model. Furthermore, Myospreader Cas9 achieves higher rates of gene editing in CRISPR reporter and Duchenne muscular dystrophy mouse models. Myospreader reveals design principles relevant to all nuclear-targeted gene therapies and highlights the importance of the spatial dimension in therapeutic development.
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