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Author Notes:

Correspondence and requests for materials should be addressed to X.G. (email: guosapphire@yahoo.com) or X.J.L. (email: xli2@emory.edu)

Z.T., W.Y., and S. Yan contributed equally to this work.

The author contributions in this manuscript were as follows: Z.T., S.L., and X.-J.L. designed the research experiments.

Z.T., W.Y., S. Yan., A.Y., X.D.L., Y.Z., J. G., J.Z. S. Yang., and Z.L. performed the experiments.

X.G. coordinated experiments. X.-J. L., S.L., W.Y., and Z.T. wrote and edited the manuscript.

We thank Dr. Shang-Hsun Yang for advice, Dr. Liangxue Lai for using his lab facilities at the Guangzhou Institutes of Biomedicine and Health, Ruxiao Xing, Xiaoyu Xue, Jianrong Liu, and other staff at Yuanxi Biotech Inc., Guangzhou for technical assistance and animal care, and Cheryl Strauss for critical reading of this manuscript.

The authors declare no competing financial interests.

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Research Funding:

The work was supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB13010300), the National Key Basic Research Program of China (2012CBA01304), the National Natural Science Foundation of China (91332206), and the State Key Laboratory of Molecular Developmental Biology, China.

Keywords:

  • Development of the nervous system
  • Functional genomics

Promoting Cas9 degradation reduces mosaic mutations in non-human primate embryos

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Journal Title:

Scientific Reports

Volume:

Volume 7

Publisher:

, Pages 42081-42081

Type of Work:

Article | Final Publisher PDF

Abstract:

CRISPR-Cas9 is a powerful new tool for genome editing, but this technique creates mosaic mutations that affect the efficiency and precision of its ability to edit the genome. Reducing mosaic mutations is particularly important for gene therapy and precision genome editing. Although the mechanisms underlying the CRSIPR/Cas9-mediated mosaic mutations remain elusive, the prolonged expression and activity of Cas9 in embryos could contribute to mosaicism in DNA mutations. Here we report that tagging Cas9 with ubiquitin-proteasomal degradation signals can facilitate the degradation of Cas9 in non-human primate embryos. Using embryo-splitting approach, we found that shortening the half-life of Cas9 in fertilized zygotes reduces mosaic mutations and increases its ability to modify genomes in non-human primate embryos. Also, injection of modified Cas9 in one-cell embryos leads to live monkeys with the targeted gene modifications. Our findings suggest that modifying Cas9 activity can be an effective strategy to enhance precision genome editing.

Copyright information:

© The Author(s) 2017.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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