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Trio deep-sequencing does not reveal unexpected off-target and on-target mutations in Cas9-edited rhesus monkeys.

ABSTRACT: 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.

SUBMITTER: Luo X

PROVIDER: S-EPMC6892871 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature

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Trio deep-sequencing does not reveal unexpected off-target and on-target mutations in Cas9-edited rhesus monkeys.

Luo Xin X He Yaoxi Y Zhang Chao C He Xiechao X Yan Lanzhen L Li Min M Hu Ting T Hu Yan Y Jiang Jin J Meng Xiaoyu X Ji Weizhi W Zhao Xudong X Zheng Ping P Xu Shuhua S Su Bing B

Nature communications 20191204 1

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 sequen ...[more]

PMID: 31797925

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