J Integr Plant Biol ›› 2021, Vol. 63 ›› Issue (9): 1595-1599.DOI: 10.1111/jipb.13089

所属专题： Biotechnology

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  • 收稿日期:2021-01-26 接受日期:2021-03-09 出版日期:2021-09-01 发布日期:2021-09-22

Efficient generation of homozygous substitutions in rice in one generation utilizing an rABE8e base editor

Chuang Wei1,2†, Chong Wang1,2†, Meng Jia1,2, Hong‐Xuan Guo1,2, Peng‐Yu Luo1,2, Mu‐Gui Wang3, Jian‐Kang Zhu3* and Hui Zhang1,2*   

  1. 1 Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai 200234, China
    2 Shanghai Key Laboratory of Plant Molecular Sciences, College of Life Sciences, Shanghai Normal University, Shanghai 200234, China
    3 Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences, The Chinese Academy of Sciences, Shanghai 200032, China

    † These authors contributed equally to this work.
    *Correspondences: Jian‐Kang Zhu (jkzhu@psc.ac.cn); Hui Zhang (zhanghui29@shnu.edu.cn, Dr. Zhang is fully responsible for the distribution of all materials associated with this article)
  • Received:2021-01-26 Accepted:2021-03-09 Online:2021-09-01 Published:2021-09-22

Abstract: A new deaminase, TadA8e, was recently evolved in the laboratory. TadA8e catalyzes DNA deamination over 1,000 times faster than ABE7.10. We developed a high-efficiency adenine base editor, rABE8e (rice ABE8e), combining monomeric TadA8e, bis-bpNLS and codon optimization. rABE8e had substantially increased editing efficiencies at NG-protospacer adjacent motif (PAM) and NGG-PAM target sequences compared with ABEmax. For most targets, rABE8e exhibited nearly 100% editing efficiency and high homozygous substitution rates in the specific editing window, especially at Positions A5 and A6. The ability to rapidly generate plant materials with homozygous base substitutions will benefit gene function research and precision molecular breeding.

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