A novel base editor SpRY-ABE8eF148A mediates efficient A-to-G base editing with a reduced off-target effect

Guo Li; Yaxian Cheng; Yeqiu Li; Hongru Ma; Zhongji Pu; Sa Li; Yiqiang Zhao; Xingxu Huang; Yuan Yao

Molecular Therapy: Nucleic Acids (Mar 2023)

A novel base editor SpRY-ABE8eF148A mediates efficient A-to-G base editing with a reduced off-target effect

Guo Li,
Yaxian Cheng,
Yeqiu Li,
Hongru Ma,
Zhongji Pu,
Sa Li,
Yiqiang Zhao,
Xingxu Huang,
Yuan Yao

Affiliations

Guo Li: ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, Zhejiang 311200, China; College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China
Yaxian Cheng: ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, Zhejiang 311200, China; College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China
Yeqiu Li: ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, Zhejiang 311200, China
Hongru Ma: ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, Zhejiang 311200, China; College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China
Zhongji Pu: ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, Zhejiang 311200, China; College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China
Sa Li: State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, China
Yiqiang Zhao: State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, China
Xingxu Huang: Zhejiang Lab, Hangzhou, Zhejiang 311121, China; School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China; CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China
Yuan Yao: ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, Zhejiang 311200, China; College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, Zhejiang 310027, China; Corresponding author Yuan Yao, ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou, Zhejiang 311200, China.

Journal volume & issue: Vol. 31
pp. 78 – 87

Abstract

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Adenine base editors (ABEs) can mediate two transition mutations, A-to-G and T-to-C, which are suitable for repairing G·C-to-T·A pathogenic variants, the most significant human pathogenic variant. By combining the protospacer adjacent motif (PAM)less SpRY nuclease with F148A-mutated TadA∗8e deaminase, we developed a new editor, SpRY-ABE8eF148A, in this study, which has narrowed the editing range and enhanced A-to-G editing efficiency in most sites with NR/YN PAMs. Furthermore, compared with SpRY-ABE8e, SpRY-ABE8eF148A significantly decreased the RNA off-target effect. Therefore, this engineered base editor, SpRY-ABE8eF148A, expanded the editing scope and improved the editing precision for G·C-to-T·A pathogenic variants. Besides, we established a bioinformatics tool, adenine base-repairing sgRNA database of pathogenic variant (ARDPM), to facilitate the development of precise editors.

Published in Molecular Therapy: Nucleic Acids

ISSN: 2162-2531 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Medicine: Therapeutics. Pharmacology
Website: https://www.cell.com/molecular-therapy-family/nucleic-acids/latest-content

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