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Development of plant cytosine base editors with the Cas12a system

文献类型： 外文期刊

作者： Wang, Huanhuan 1 ; Liang, Jing 1 ; Chen, Like 1 ; Deng, Bufang 1 ; Gu, Dongfang 2 ; Liu, Xiaoshuang 1 ; Jin, Shan 2 ; Xu, Rongfang 2 ; Qin, Ruiying 2 ; Zhu, Yitong 1 ; Zhao, Liangxia 1 ; Kou, Dourong 2 ; Chen, Yanjun 1 ; Jiang, Yingli 1 ; Li, Juan 2 ; Wei, Pengcheng 1 ;

作者机构： 1.Anhui Agr Univ, Coll Agron, Hefei 230036, Anhui, Peoples R China

2.Anhui Acad Agr Sci, Rice Res Inst, Key Lab Rice Genet Breeding Anhui Prov, Hefei 230031, Anhui, Peoples R China

关键词： Cas12a; Base editing; Rice; ABE; CBE

期刊名称：CROP JOURNAL （ 影响因子：6.6； 五年影响因子：6.5 ）

ISSN： 2095-5421

年卷期： 2023 年 11 卷 5 期

页码：

收录情况： SCI

摘要： 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. (c) 2023 Crop Science Society of China and Institute of Crop Science, CAAS. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. 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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