Harnessing A3G for efficient and selective C-to-T conversion at C-rich sequences
Yu, Wenxia1,2; Li, Jianan1,2,3; Huang, Shisheng1,2; Li, Xiangyang1,2; Li, Ping4; Li, Guanglei1; Liang, Aibin4; Chi, Tian1,5; Huang, Xingxu1,6
2021-02
Source PublicationBMC BIOLOGY
ISSN1741-7007
EISSN1741-7007
Volume19Issue:1Pages:#VALUE!
Status已发表
DOI10.1186/s12915-020-00879-0
Abstract

Background Site-specific C>T DNA base editing has been achieved by recruiting cytidine deaminases to the target C using catalytically impaired Cas proteins; the target C is typically located within 5-nt editing window specified by the guide RNAs. The prototypical cytidine base editor BE3, comprising rat APOBEC1 (rA1) fused to nCas9, can indiscriminately deaminate multiple C's within the editing window and also create substantial off-target edits on the transcriptome. A powerful countermeasure for the DNA off-target editing is to replace rA1 with APOBEC proteins which selectively edit C's in the context of specific motifs, as illustrated in eA3A-BE3 which targets TC. However, analogous editors selective for other motifs have not been described. In particular, it has been challenging to target a particular C in C-rich sequences. Here, we sought to confront this challenge and also to overcome the RNA off-target effects seen in BE3. Results By replacing rA1 with an optimized human A3G (oA3G), we developed oA3G-BE3, which selectively targets CC and CCC and is also free of global off-target effects on the transcriptome. Furthermore, we created oA3G-BE4max, an upgraded version of oA3G-BE3 with robust on-target editing. Finally, we showed that oA3G-BE4max has negligible Cas9-independent off-target effects at the genome. Conclusions oA3G-BE4max can edit C(C)C with high efficiency and selectivity, which complements eA3A-editors to broaden the collective editing scope of motif selective editors, thus filling a void in the base editing tool box.

KeywordBase editing Apobec 3G Motif C-rich
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Indexed BySCI
Language英语
WOS Research AreaLife Sciences & Biomedicine - Other Topics
WOS SubjectBiology
WOS IDWOS:000619418300001
PublisherBMC
Original Document TypeArticle
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Cited Times [WOS]:0   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttps://kms.shanghaitech.edu.cn/handle/2MSLDSTB/125792
Collection生命科学与技术学院_硕士生
生命科学与技术学院_PI研究组_池天组
生命科学与技术学院_PI研究组_黄行许组
生命科学与技术学院_博士生
Co-First AuthorLi, Jianan
Affiliation1.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China;
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China;
3.Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China;
4.Tongji Univ, Dept Hematol, Tongji Hosp, Shanghai 200092, Peoples R China;
5.Yale Univ, Sch Med, Dept Immunobiol, New Haven, CT USA;
6.Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, CAS Ctr Excellence Mol Cell Sci, Shanghai 200031, Peoples R China
First Author AffilicationSchool of Life Science and Technology
First Signature AffilicationSchool of Life Science and Technology
Recommended Citation
GB/T 7714
Yu, Wenxia,Li, Jianan,Huang, Shisheng,et al. Harnessing A3G for efficient and selective C-to-T conversion at C-rich sequences[J]. BMC BIOLOGY,2021,19(1):#VALUE!.
APA Yu, Wenxia.,Li, Jianan.,Huang, Shisheng.,Li, Xiangyang.,Li, Ping.,...&Huang, Xingxu.(2021).Harnessing A3G for efficient and selective C-to-T conversion at C-rich sequences.BMC BIOLOGY,19(1),#VALUE!.
MLA Yu, Wenxia,et al."Harnessing A3G for efficient and selective C-to-T conversion at C-rich sequences".BMC BIOLOGY 19.1(2021):#VALUE!.
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