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Expanded targeting scope and enhanced base editing efficiency in rabbit using optimized xCas9(3.7)

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Abstract

Evolved xCas9(3.7) variant with broad PAM compatibility has been reported in cell lines, while its editing efficiency was site-specific. Here, we show that xCas9(3.7) can recognize a broad PAMs including NGG, NGA, and NGT, in both embryos and Founder (F0) rabbits. Furthermore, the codon-optimized xCas9-derived base editors, exBE4 and exABE, can dramatically improve the base editing efficiencies in rabbit embryos. Our results demonstrated that the optimized xCas9 with expanded PAM compatibility and enhanced base editing efficiency could be used for precise gene modifications in organisms.

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The authors state that all data necessary for confirming the conclusions presented in the article are represented fully within the article or from the authors upon request.

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Acknowledgements

The authors would like to thank Peiran Hu for assistance at the Embryo Engineering Center for the critical technical assistance.

Funding

This study was financially supported by the National Key Research and Development Program of China Stem Cell and Translational Research (2017YFA0105101), The Program for Changjiang Scholars and Innovative Research Team in University (No.IRT_16R32), The Strategic Priority Research Program of the Chinese Academy of Sciences (XDA16030501, XDA16030503), and Key Research & Development Program of Guangzhou Regenerative Medicine and Health Guangdong Laboratory(2018GZR110104004).

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Author notes

  1. Zhiquan Liu, Mao Chen, Huanhuan Shan, and Siyu Chen have contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Animal Science, Jilin University, Changchun, 130062, China

    Zhiquan Liu, Mao Chen, Huanhuan Shan, Siyu Chen, Yuxin Xu, Yuning Song, Hongming Yuan, Hongsheng Ouyang, Zhanjun Li & Liangxue Lai

  2. CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China

    Quanjun Zhang & Liangxue Lai

  3. Guangzhou Regenerative Medicine and Health GuangDong Laboratory (GRMH-GDL), Guangzhou, 510005, China

    Liangxue Lai

  4. Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, 100101, China

    Liangxue Lai

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  1. Zhiquan Liu
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Contributions

ZL, MC, HS, and SC performed the experiment; ZL and LL conceived the idea and provided funding support; ZL, ZL, and LL wrote the manuscript. All authors reviewed the manuscript.

Corresponding authors

Correspondence to Zhanjun Li or Liangxue Lai.

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Liu, Z., Chen, M., Shan, H. et al. Expanded targeting scope and enhanced base editing efficiency in rabbit using optimized xCas9(3.7). Cell. Mol. Life Sci. 76, 4155–4164 (2019). https://doi.org/10.1007/s00018-019-03110-8

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