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

Author

Listed:
  • Xin Luo

    (Chinese Academy of Sciences
    University of Chinese Academy of Sciences
    Chinese Academy of Sciences)

  • Yaoxi He

    (Chinese Academy of Sciences
    University of Chinese Academy of Sciences
    Chinese Academy of Sciences)

  • Chao Zhang

    (Shanghai Institutes for Biological Sciences, CAS)

  • Xiechao He

    (Chinese Academy of Sciences
    Chinese Academy of Sciences)

  • Lanzhen Yan

    (Chinese Academy of Sciences
    Chinese Academy of Sciences)

  • Min Li

    (Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Ting Hu

    (Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Yan Hu

    (Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Jin Jiang

    (Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Xiaoyu Meng

    (Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Weizhi Ji

    (Kunming University of Science and Technology)

  • Xudong Zhao

    (Chinese Academy of Sciences
    Chinese Academy of Sciences)

  • Ping Zheng

    (Chinese Academy of Sciences
    Chinese Academy of Sciences)

  • Shuhua Xu

    (Shanghai Institutes for Biological Sciences, CAS
    Chinese Academy of Sciences
    ShanghaiTech University
    Collaborative Innovation Centre of Genetics and Development)

  • Bing Su

    (Chinese Academy of Sciences
    Chinese Academy of Sciences
    Chinese Academy of Sciences)

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.

Suggested Citation

  • 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 & Bing Su, 2019. "Trio deep-sequencing does not reveal unexpected off-target and on-target mutations in Cas9-edited rhesus monkeys," Nature Communications, Nature, vol. 10(1), pages 1-7, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-13481-y
    DOI: 10.1038/s41467-019-13481-y
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    Cited by:

    1. Ida Höijer & Anastasia Emmanouilidou & Rebecka Östlund & Robin Schendel & Selma Bozorgpana & Marcel Tijsterman & Lars Feuk & Ulf Gyllensten & Marcel Hoed & Adam Ameur, 2022. "CRISPR-Cas9 induces large structural variants at on-target and off-target sites in vivo that segregate across generations," Nature Communications, Nature, vol. 13(1), pages 1-10, December.

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