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Efficient and heritable A-to-K base editing in rice and tomato

文献类型： 外文期刊

作者： Li, Xinbo 1 ; Xie, Jiyong 4 ; Dong, Chao 1 ; Zheng, Zai 1 ; Shen, Rundong 1 ; Cao, Xuesong 6 ; Chen, Xiaoyan 1 ; Wang, Mugui 1 ; Zhu, Jian-Kang 1 ; Tian, Yifu 1 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Crop Sci, Key Lab Gene Editing Technol Hainan, Minist Agr & Rural Affairs, Sanya 572024, Hainan, Peoples R China

2.Chinese Acad Agr Sci, Natl Nanfan Res Inst, Sanya 572024, Hainan, Peoples R China

3.Hainan Yazhou Bay Seed Lab, Sanya 572024, Hainan, Peoples R China

4.Chinese Acad Sci, Shanghai Ctr Plant Stress Biol, CAS Ctr Excellence Mol Plant Sci, Shanghai 201602, Peoples R China

5.Univ Chinese Acad Sci, Beijing 100049, Peoples R China

6.Southern Univ Sci & Technol, Inst Adv Biotechnol, Shenzhen 518055, Peoples R China

7.Southern Univ Sci & Technol, Sch Life Sci, Shenzhen 518055, Peoples R China

期刊名称：HORTICULTURE RESEARCH （ 影响因子：8.7； 五年影响因子：9.0 ）

ISSN： 2662-6810

年卷期： 2024 年 11 卷 1 期

页码：

收录情况： SCI

摘要： Cytosine and adenosine base editors (CBE and ABE) have been widely used in plants, greatly accelerating gene function research and crop breeding. Current base editors can achieve efficient A-to-G and C-to-T/G/A editing. However, efficient and heritable A-to-Y (A-to-T/C) editing remains to be developed in plants. In this study, a series of A-to-K base editor (AKBE) systems were constructed for monocot and dicot plants. Furthermore, nSpCas9 was replaced with the PAM-less Cas9 variant (nSpRY) to expand the target range of the AKBEs. Analysis of 228 T0 rice plants and 121 T0 tomato plants edited using AKBEs at 18 endogenous loci revealed that, in addition to highly efficient A-to-G substitution (41.0% on average), the plant AKBEs can achieve A-to-T conversion with efficiencies of up to 25.9 and 10.5% in rice and tomato, respectively. Moreover, the rice-optimized AKBE generates A-to-C conversion in rice, with an average efficiency of 1.8%, revealing the significant value of plant-optimized AKBE in creating genetic diversity. Although most of the A-to-T and A-to-C edits were chimeric, desired editing types could be transmitted to the T1 offspring, similar to the edits generated by the traditional ABE8e. Besides, using AKBEs to target tyrosine (Y, TAT) or cysteine (C, TGT) achieved the introduction of an early stop codon (TAG/TAA/TGA) of target genes, demonstrating its potential use in gene disruption.

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