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

Research output: Contribution to journalArticlepeer-review

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 languageEnglish (US)
Pages (from-to)728-740.e6
JournalMolecular Cell
Volume79
Issue number5
DOIs
StatePublished - 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.

Keywords

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

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't
  • Validation Study

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