Rationally Designed APOBEC3B Cytosine Base Editors with Improved Specificity

Shuai Jin; Hongyuan Fei; Zixu Zhu; Yingfeng Luo; Jinxing Liu; Shenghan Gao; Feng Zhang; Yu Hang Chen; Yanpeng Wang; Caixia Gao

doi:10.1016/j.molcel.2020.07.005

Rationally Designed APOBEC3B Cytosine Base Editors with Improved Specificity

Shuai Jin, Hongyuan Fei, Zixu Zhu, Yingfeng Luo, Jinxing Liu, Shenghan Gao, Feng Zhang, Yu Hang Chen, Yanpeng Wang, Caixia Gao

Plant and Microbial Biology

Research output: Contribution to journal › Article › peer-review

96 Scopus citations

Abstract

Jin et al. establish nSaCas9-mediated orthogonal R-loop assay, a rapid, high-throughput, and inexpensive method for assessing CBEs in plants. Using it, they assess the specificity of 25 rationally designed A3Bctd-BE3s, identifying A3Bctd-VHM-BE3 and A3Bctd-KKR-BE3 with high specificity and precision.

Original language	English (US)
Pages (from-to)	728-740.e6
Journal	Molecular Cell
Volume	79
Issue number	5
DOIs	https://doi.org/10.1016/j.molcel.2020.07.005
State	Published - Sep 3 2020

Bibliographical note

Funding Information:
We thank Dr. Fei Lu (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences [IGDB, CAS]) for advice in the statistics analysis. This work was supported by grants from the National Transgenic Science and Technology Program (2019ZX08010-003 and 2019ZX08010-001), the Strategic Priority Research Program of the CAS (Precision Seed Design and Breeding, XDA24020102), the National Natural Science Foundation of China (31788103), the Youth Innovation Promotion Association of the Chinese Academy of Sciences (2020000003), and by a grant from the United Soybean Board (1920-152-0131-B) to F.Z. Y.W. and C.G. designed the experiments. S.J. H.F. and Z.Z. performed most of the experiments. Y.L. S.J. and S.G. performed the WGS analysis. J.L. performed the rice transformation experiments. Y.-H.C. analyzed the structure of the A3Bctd protein. Y.W. and C.G. supervised the project. S.J. H.F. Z.Z. Y.W. F.Z. and C.G. wrote the manuscript. The authors have submitted two patent applications based on the results reported in this article.

Funding Information:
We thank Dr. Fei Lu (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences [IGDB, CAS]) for advice in the statistics analysis. This work was supported by grants from the National Transgenic Science and Technology Program ( 2019ZX08010-003 and 2019ZX08010-001 ), the Strategic Priority Research Program of the CAS (Precision Seed Design and Breeding, XDA24020102 ), the National Natural Science Foundation of China ( 31788103 ), the Youth Innovation Promotion Association of the Chinese Academy of Sciences (2020000003), and by a grant from the United Soybean Board ( 1920-152-0131-B ) to F.Z.

Publisher Copyright:
© 2020 Elsevier Inc.

Keywords

A3Bctd deaminase
cytosine base editors
nSaCas9
off-target editing
rationally designing
specificity
whole-genome sequencing

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title = "Rationally Designed APOBEC3B Cytosine Base Editors with Improved Specificity",

abstract = "Jin et al. establish nSaCas9-mediated orthogonal R-loop assay, a rapid, high-throughput, and inexpensive method for assessing CBEs in plants. Using it, they assess the specificity of 25 rationally designed A3Bctd-BE3s, identifying A3Bctd-VHM-BE3 and A3Bctd-KKR-BE3 with high specificity and precision.",

keywords = "A3Bctd deaminase, cytosine base editors, nSaCas9, off-target editing, rationally designing, specificity, whole-genome sequencing",

author = "Shuai Jin and Hongyuan Fei and Zixu Zhu and Yingfeng Luo and Jinxing Liu and Shenghan Gao and Feng Zhang and Chen, {Yu Hang} and Yanpeng Wang and Caixia Gao",

note = "Funding Information: We thank Dr. Fei Lu (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences [IGDB, CAS]) for advice in the statistics analysis. This work was supported by grants from the National Transgenic Science and Technology Program (2019ZX08010-003 and 2019ZX08010-001), the Strategic Priority Research Program of the CAS (Precision Seed Design and Breeding, XDA24020102), the National Natural Science Foundation of China (31788103), the Youth Innovation Promotion Association of the Chinese Academy of Sciences (2020000003), and by a grant from the United Soybean Board (1920-152-0131-B) to F.Z. Y.W. and C.G. designed the experiments. S.J. H.F. and Z.Z. performed most of the experiments. Y.L. S.J. and S.G. performed the WGS analysis. J.L. performed the rice transformation experiments. Y.-H.C. analyzed the structure of the A3Bctd protein. Y.W. and C.G. supervised the project. S.J. H.F. Z.Z. Y.W. F.Z. and C.G. wrote the manuscript. The authors have submitted two patent applications based on the results reported in this article. Funding Information: We thank Dr. Fei Lu (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences [IGDB, CAS]) for advice in the statistics analysis. This work was supported by grants from the National Transgenic Science and Technology Program ( 2019ZX08010-003 and 2019ZX08010-001 ), the Strategic Priority Research Program of the CAS (Precision Seed Design and Breeding, XDA24020102 ), the National Natural Science Foundation of China ( 31788103 ), the Youth Innovation Promotion Association of the Chinese Academy of Sciences (2020000003), and by a grant from the United Soybean Board ( 1920-152-0131-B ) to F.Z. Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",

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AU - Zhu, Zixu

AU - Luo, Yingfeng

AU - Liu, Jinxing

AU - Gao, Shenghan

AU - Zhang, Feng

AU - Chen, Yu Hang

AU - Wang, Yanpeng

AU - Gao, Caixia

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ER -

Rationally Designed APOBEC3B Cytosine Base Editors with Improved Specificity

Abstract

Bibliographical note

Keywords

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