Trio deep-sequencing does not reveal unexpected off-target and on-target mutations in Cas9-edited rhesus monkeys
Trio deep-sequencing does not reveal unexpected off-target and on-target mutations in Cas9-edited rhesus monkeys
复制标题
Trio 深度测序并未揭示 Cas9 编辑的恒河猴中意外的脱靶和在靶突变
DOI：
10.1038/s41467-019-13481-y
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发表时间：
2019-12-04
影响因子：
16.6
通讯作者：
Su, Bing
中科院分区：
文献类型：
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作者：
Luo, Xin;He, Yaoxi;Su, Bing
CRISPR-Cas9是一种广泛使用的基因组编辑工具，但其脱靶效应和靶向复杂突变仍然是一个问题，特别是考虑到未来的临床应用。非人灵长类动物（NHP）与人类具有密切的遗传和生理相似性，使其成为开发基于Cas9疗法的理想临床前模型。然而，据我们所知，尚未在NHP中进行全面的体内脱靶和中靶评估。在这里，我们对Cas9处理的恒河猴进行全基因组三重测序。我们只发现了少量可以用预期的自发突变解释的从头突变，没有检测到非预期的脱靶突变（OTM）。此外，长读段测序数据未检测到靶区域中的大结构变体。
CRISPR-Cas9 is a widely-used genome editing tool, but its off-target effect and on-target complex mutations remain a concern, especially in view of future clinical applications. Non-human primates (NHPs) share close genetic and physiological similarities with humans, making them an ideal preclinical model for developing Cas9-based therapies. However, to our knowledge no comprehensive in vivo off-target and on-target assessment has been conducted in NHPs. Here, we perform whole genome trio sequencing of Cas9-treated rhesus monkeys. We only find a small number of de novo mutations that can be explained by expected spontaneous mutations, and no unexpected off-target mutations (OTMs) were detected. Furthermore, the long-read sequencing data does not detect large structural variants in the target region.