Improving adenine and dual base editors through introduction of TadA-8e and Rad51DBD

Niannian Xue; Xu Liu; Dan Zhang; Youming Wu; Yi Zhong; Jinxin Wang; Wenjing Fan; Haixia Jiang; Biyun Zhu; Xiyu Ge; Rachel V L Gonzalez; Liang Chen; Shun Zhang; Peilu She; Zhilin Zhong; Jianjian Sun; Xi Chen; Liren Wang; Zhimin Gu; Ping Zhu; Mingyao Liu; Dali Li; Tao P Zhong; Xiaohui Zhang

doi:10.1038/s41467-023-36887-1

Improving adenine and dual base editors through introduction of TadA-8e and Rad51DBD

Nat Commun. 2023 Mar 3;14(1):1224. doi: 10.1038/s41467-023-36887-1.

Authors

Niannian Xue^#¹, Xu Liu^#¹, Dan Zhang^#¹, Youming Wu^#^{2

3}, Yi Zhong¹, Jinxin Wang^{2

3}, Wenjing Fan^{2

3}, Haixia Jiang⁴, Biyun Zhu¹, Xiyu Ge⁵, Rachel V L Gonzalez⁶, Liang Chen¹, Shun Zhang¹, Peilu She^{1

7}, Zhilin Zhong¹, Jianjian Sun^{1

7}, Xi Chen⁸, Liren Wang¹, Zhimin Gu^{2

3}, Ping Zhu⁷, Mingyao Liu^{1

8}, Dali Li⁹, Tao P Zhong¹⁰, Xiaohui Zhang^{11

12

13}

Affiliations

¹ Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.
² Institute of Systems Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
³ Suzhou Institute of Systems Medicine, Suzhou, China.
⁴ School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai, China.
⁵ Department of Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
⁶ Department of Physiology and Cellular Biophysics, Columbia University, Manhattan, NY, USA.
⁷ Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.
⁸ BRL Medicine, Inc., Shanghai, China.
⁹ Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China. dlli@bio.ecnu.edu.cn.
¹⁰ Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China. tzhong@bio.ecnu.edu.cn.
¹¹ Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China. zhangxiaohui3040@126.com.
¹² Institute of Systems Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. zhangxiaohui3040@126.com.
¹³ Suzhou Institute of Systems Medicine, Suzhou, China. zhangxiaohui3040@126.com.

^# Contributed equally.

Abstract

Base editors, including dual base editors, are innovative techniques for efficient base conversions in genomic DNA. However, the low efficiency of A-to-G base conversion at positions proximal to the protospacer adjacent motif (PAM) and the A/C simultaneous conversion of the dual base editor hinder their broad applications. In this study, through fusion of ABE8e with Rad51 DNA-binding domain, we generate a hyperactive ABE (hyABE) which offers improved A-to-G editing efficiency at the region (A₁₀-A₁₅) proximal to the PAM, with 1.2- to 7-fold improvement compared to ABE8e. Similarly, we develop optimized dual base editors (eA&C-BEmax and hyA&C-BEmax) with markedly improved simultaneous A/C conversion efficiency (1.2-fold and 1.5-fold improvement, respectively) compared to A&C-BEmax in human cells. Moreover, these optimized base editors catalyze efficiently nucleotide conversions in zebrafish embryos to mirror human syndrome or in human cells to potentially treat genetic diseases, indicating their great potential in broad applications for disease modeling and gene therapy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenine*
Animals
Catalysis
Genetic Therapy
Humans
Nucleotides
Zebrafish*

Substances

Adenine
Nucleotides