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

Xin Luo, Yaoxi He, Chao Zhang, Xiechao He, Lanzhen Yan, Min Li, Ting Hu, Yan Hu, Jin Jiang, Xiaoyu Meng, Weizhi Ji, Xudong Zhao, Ping Zheng (), Shuhua Xu () and Bing Su ()
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Xin Luo: Chinese Academy of Sciences
Yaoxi He: Chinese Academy of Sciences
Chao Zhang: Shanghai Institutes for Biological Sciences, CAS
Xiechao He: Chinese Academy of Sciences
Lanzhen Yan: Chinese Academy of Sciences
Min Li: Chinese Academy of Sciences
Ting Hu: Chinese Academy of Sciences
Yan Hu: Chinese Academy of Sciences
Jin Jiang: Chinese Academy of Sciences
Xiaoyu Meng: Chinese Academy of Sciences
Weizhi Ji: Kunming University of Science and Technology
Xudong Zhao: Chinese Academy of Sciences
Ping Zheng: Chinese Academy of Sciences
Shuhua Xu: Shanghai Institutes for Biological Sciences, CAS
Bing Su: Chinese Academy of Sciences

Nature Communications, 2019, vol. 10, issue 1, 1-7

Abstract: 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.

Date: 2019
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DOI: 10.1038/s41467-019-13481-y

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