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Diverse nucleotide substitutions in rice base editing mediated by novel TadA variants

文献类型： 外文期刊

作者： Yu, Man 1 ; Kuang, Yongjie 1 ; Wang, Chenyang 1 ; Wu, Xuemei 1 ; Li, Shaofang 5 ; Zhang, Dawei 4 ; Sun, Wenxian 2 ; Zhou, Xueping 1 ; Ren, Bin 1 ; Zhou, Huanbin 1 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China

2.China Agr Univ, Dept Plant Pathol, Beijing 100193, Peoples R China

3.Minist Agr & Rural Affairs, Sci Observing & Expt Stn Crop Pests Guilin, Guilin 541399, Peoples R China

4.Sichuan Univ, Coll Life Sci, Minist Educ, Key Lab Bioresource & Ecoenvironm,State Key Lab Hy, Chengdu 610064, Peoples R China

5.Beijing Acad Agr & Forestry Sci, Natl Engn Res Ctr Vegetables, Beijing Vegetable Res Ctr, State Key Lab Vegetable Biobreeding, Beijing 100097, Peoples R China

6.Zhejiang Univ, Inst Biotechnol, State Key Lab Rice Biol, Hangzhou 310058, Zhejiang, Peoples R China

7.Minist Agr & Rural Affairs, Key Lab Gene Editing Technol Hainan, Sanya 572024, Peoples R China

关键词： CRISPR; TadA variants; cytosine base editing; dual base editor; rice

期刊名称：PLANT COMMUNICATIONS （ 影响因子：9.4； 五年影响因子：9.4 ）

ISSN： 2590-3462

年卷期： 2024 年 5 卷 8 期

页码：

收录情况： SCI

摘要： CRISPR-mediated base editors have been widely used to correct defective alleles and create novel alleles by artificial evolution for the rapid genetic improvement of crops. The editing capabilities of base editors strictly rely on the performance of various nucleotide modification enzymes. Compared with the well- developed adenine base editors (ABEs), cytosine base editors (CBEs) and dual base editors suffer from unstable editing efficiency and patterns at different genomic loci in rice, significantly limiting their application. Here, we comprehensively examined the base editing activities of multiple evolved TadA8e variants in rice. We found that both TadA-CDd and TadA-E27R/N46L achieved more robust C-to-T editing than previously reported hyperactive hAID*D, D , and TadA-CDd outperformed TadA-E27R/N46L. A C-to-G base editor (CGBE) engineered with TadA-CDd and OsUNG performed highly efficient C-to-G editing in rice compared with that of TadA-N46P. In addition, a dual base editor constructed with a single protein, TadDE, enabled simultaneous, highly efficient C-to-T and A-to-G editing in rice. Collectively, our results demonstrate that TadA8e derivatives improve both CBEs and dual base editors in rice, providing a powerful way to induce diverse nucleotide substitutions for plant genome editing.

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