期刊
NATURE METHODS
卷 17, 期 6, 页码 600-+出版社
NATURE PORTFOLIO
DOI: 10.1038/s41592-020-0832-x
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资金
- R&D Program of China [2018YFC2000100, 2017YFC1001302, 2017YFC0909701]
- CAS Strategic Priority Research Program [XDB32060000, XDBS01060100]
- National Natural Science Foundation of China [31871502, 31522037, 31822035, 31922048, 31925016, 91957122]
- Basic Frontier Scientific Research Program of Chinese Academy of Sciences [ZDBS-LY-SM001]
- Shanghai Municipal Science and Technology Major Project [2018SHZDZX05]
- Shanghai City Committee of science and technology project [18411953700, 18JC1410100, 16JC1420202]
- National Science and Technology Major Project [2015ZX10004801-005]
- National Key Research and Development Program of China [2017YFA0505500, 2016YFC0901704]
- Agricultural Science and Technology Innovation Program
- International Partnership Program of Chinese Academy of Sciences [153D31KYSB20170059]
Cytosine base editors (CBEs) offer a powerful tool for correcting point mutations, yet their DNA and RNA off-target activities have caused concerns in biomedical applications. We describe screens of 23 rationally engineered CBE variants, which reveal mutation residues in the predicted DNA-binding site can dramatically decrease the Cas9-independent off-target effects. Furthermore, we obtained a CBE variant-YE1-BE3-FNLS-that retains high on-target editing efficiency while causing extremely low off-target edits and bystander edits. Structural and biochemical insights help engineer a cytosine base editor variant that possesses improved on-target activity with minimal DNA and RNA off-target editing.
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