J Integr Plant Biol.

• Breakthrough Report •    

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. 1Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai 200234, China.
    2Shanghai Key Laboratory of Plant Molecular Sciences, College of Life Sciences, Shanghai Normal University, 100 Guilin Road, Shanghai 200234, China.
    3Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai 200032, China.

    These authors contributed equally to this work.
    *Correspondence: Hui Zhang (zhanghui29@shnu.edu.cn, Dr. Zhang is fully responsible for the distribution of all materials associated with this article); Jian-kang Zhu (jkzhu@psc.ac.cn).
  • Received:2021-01-26 Accepted:2021-03-09 Online:2021-03-10

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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