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Research paper | Open Access

Development of plant cytosine base editors with the Cas12a system

Huanhuan Wanga,1, Jing Lianga,1, Like Chena, Bufang Denga, Dongfang Gub, Xiaoshuang Liua, Shan Jinb, Rongfang Xub, Ruiying Qinb, Yitong Zhua, Liangxia Zhaoa, Dourong Koub, Yanjun Chena, Yingli Jianga, Juan Lib( ), Pengcheng Weia( )
College of Agronomy, Anhui Agricultural University, Hefei 230036, Anhui, China
Key Laboratory of Rice Genetic Breeding of Anhui Province, Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei 230031, Anhui, China

1 These authors contributed equally to this work.

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Abstract

Base editors of the Cas9 system have been widely used for precise nucleotide substitution in crops. In this study, Cas12a was applied to construct plant cytosine base editors (CBEs). The main elements of Cas12a-CBEs were engineered and their efficiency was evaluated in stably transformed rice cells. An optimized ttCas12a-hyA3Bctd editor, consisting of a LbCas12a variant carrying catalytic inactive D832A and temperature-tolerance D156R double mutations, a truncated human APOBEC3B deaminase, a human RAD51 single-stranded DNA-binding domain, and double copies of UGI, outperformed other Cas12a-CBEs in base editing efficiency. In T0 transgenic rice plants, ttCas12a-hyA3Bctd edited an average of 42.01% and a maximum of 68.75% of lines at six genomic targets. A-to-G conversions were generated in rice by an adenine base editor with a similar architecture to the optimized CBE. Our results provide preliminary evidence for the feasibility of robust and efficient plant Cas12a base editing systems, which could be useful for precise crop breeding.

Keywords

RiceBase editingCas12aABECBE

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The Crop Journal
Volume 11 Issue 5,
October 2023
Pages 1451-1457
DOI: 10.1016/j.cj.2023.03.002
Cite this article:
Wang H, Liang J, Chen L, et al. Development of plant cytosine base editors with the Cas12a system. The Crop Journal, 2023, 11(5): 1451-1457. https://doi.org/10.1016/j.cj.2023.03.002

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Received: 03 November 2022
Revised: 09 February 2023
Accepted: 05 March 2023
Published: 27 March 2023
© 2023 Crop Science Society of China and Institute of Crop Science, CAAS.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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