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Improving plant C-to-G base editors with cold-adapted glycosylase and TadA-8e variants

文献类型： 外文期刊

作者： Jiang, Yingli 1 ; Xiao, Zhi 1 ; Luo, Zhaopeng 4 ; Zhou, Suhuai 1 ; Tong, Chaoyun 1 ; Jin, Shan 1 ; Liu, Xiaoshuang 1 ; Qin, Ruiying 2 ; Xu, Rongfang 2 ; Pan, Lang 5 ; Li, Juan 2 ; Wei, Pengcheng 1 ;

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

2.Anhui Acad Agr Sci, Anhui Prov Key Lab Rice Germplasm Innovat & Mol, Hefei 230031, Peoples R China

3.Anhui Agr Univ, Adv Acad, Res Ctr Biol Breeding Technol, Hefei 230036, Peoples R China

4.Zhengzhou Tobacco Res Inst CNTC, China Tobacco Gene Res Ctr, Zhengzhou 450001, Peoples R China

5.Hunan Agr Univ, Coll Plant Protect, Changsha 410128, Peoples R China

期刊名称：TRENDS IN BIOTECHNOLOGY （ 影响因子：14.9； 五年影响因子：16.6 ）

ISSN： 0167-7799

年卷期： 2025 年 43 卷 7 期

页码：

收录情况： SCI

摘要： Plant cytosine (C)-to-guanine (G) base editors (CGBEs) have been established but suffer from limited editing efficiencies and low outcome purities. This study engineered a cod uracil DNA glycosylase (cod UNG, coUNG) from the cold-adapted fish Gadus morhua for plant CGBE, demonstrating 1.71-to 2.54-fold in-creases in C-to-G editing efficiency compared with the CGBE using human UNG (HUNG). Further engineering took advantage of TadA-8e-derived cytidine deaminases (TadA-CDs). These variants induced C substitutions with efficiencies ranging from 26.28% to 30.82% in rice cells, whereas adenine (A) conversion was negligible. By integrating coUNG and TadA-CDc elements with SpCas9 nickase, the resulting CDC-CGBEco achieved pure C-to-G editing without byproducts in up to 52.08% of transgenic lines. Whole-genome sequencing (WGS) analysis revealed no significant off-target effects of the CDC-BEs in rice. In addition, CDC-CGBEco enabled precise C-to-G editing in soybean and to-bacco. These engineered CGBEs enhanced editing efficiency, purity, and specificity, suggesting their broad potential for applications in scientific research and crop breeding.

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