TadA orthologs enable both cytosine and adenine editing of base editors

被引：17

作者：

Zhang, Shuqian ^{[1
,2
]}

Yuan, Bo ^{[3
]}

Cao, Jixin ^{[4
]}

Song, Liting ^{[4
]}

Chen, Jinlong ^{[1
]}

Qiu, Jiayi ^{[1
]}

Qiu, Zilong ^{[3
,5
,6
]}

Zhao, Xing-Ming ^{[4
,7
,8
]}

Chen, Jingqi ^{[4
,7
,8
]}

Cheng, Tian-Lin ^{[1
]}

机构：

[1] Fudan Univ, Childrens Hosp, Inst Translat Brain Res, Inst Pediat,State Key Lab Med Neurobiol,MOE Front, Shanghai, Peoples R China

[2] Shandong Univ, Dept Pediat, Qilu Hosp, Jinan 250012, Peoples R China

[3] Chinese Acad Sci, Inst Neurosci, CAS Ctr Excellence Brain Sci & Intelligence Techno, State Key Lab Neurosci, Shanghai 200031, Peoples R China

[4] Fudan Univ, Inst Sci & Technol Brain Inspired Intelligence, Shanghai, Peoples R China

[5] Fudan Univ, Natl Clin Res Ctr Aging & Med, Huashan Hopsital, Shanghai 200032, Peoples R China

[6] Shanghai Jiao Tong Univ, Songjiang Hosp, Songjiang Inst, Sch Med, Shanghai, Peoples R China

[7] Fudan Univ, Inst Brain Sci, State Key Lab Med Neurobiol, Shanghai, Peoples R China

[8] Fudan Univ, MOE Frontiers Ctr Brain Sci, MOE Key Lab Computat Neurosci & Brain Inspired Int, Shanghai, Peoples R China

来源：

NATURE COMMUNICATIONS | 2023年 / 14卷 / 01期

基金：

上海市自然科学基金; 中国国家自然科学基金; 国家重点研发计划;

关键词：

GENOMIC DNA; OFF-TARGET; RNA; INOSINE;

D O I：

10.1038/s41467-023-36003-3

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Properties of cytidine and adenosine deaminases lead to off-target effects for cytosine base editors (CBEs) and adenine base editors (ABEs). Here the authors report that 25 TadA orthologs could be engineered to generate functional ABEs, CBEs or ACBEs via single/double mutations with minimised off-targets. Cytidine and adenosine deaminases are required for cytosine and adenine editing of base editors respectively, and no single deaminase could enable concurrent and comparable cytosine and adenine editing. Additionally, distinct properties of cytidine and adenosine deaminases lead to various types of off-target effects, including Cas9-indendepent DNA off-target effects for cytosine base editors (CBEs) and RNA off-target effects particularly severe for adenine base editors (ABEs). Here we demonstrate that 25 TadA orthologs could be engineered to generate functional ABEs, CBEs or ACBEs via single or double mutations, which display minimized Cas9-independent DNA off-target effects and genotoxicity, with orthologs B5ZCW4, Q57LE3, E8WVH3, Q13XZ4 and B3PCY2 as promising candidates for further engineering. Furthermore, RNA off-target effects of TadA ortholog-derived base editors could be further reduced or even eliminated by additional single mutation. Taken together, our work expands the base editing toolkits, and also provides important clues for the potential evolutionary process of deaminases.

引用

页数：10

共 42 条

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