Highly efficient base editing in Staphylococcus aureus using an engineered CRISPR RNA-guided cytidine deaminase

Chemical Science
Tongnian GuQuanjiang Ji

Abstract

Novel therapeutic means against Staphylococcus aureus infections are urgently needed due to the emergence of drug-resistant S. aureus. We report the development of a CRISPR RNA-guided cytidine deaminase (pnCasSA-BEC), enabling highly efficient gene inactivation and point mutations in S. aureus. We engineered a fusion of a Cas9 nickase (Cas9D10A) and a cytidine deaminase (APOBEC1) that can be guided to a target genomic locus for gene inactivation via generating a premature stop codon. The pnCasSA-BEC system nicks the non-edited strand of the genomic DNA, directly catalyzes the conversion of cytidine (C) to uridine (U), and relies on DNA replication to achieve C → T (G → A) conversion without using donor repair templates. The development of the base-editing system will dramatically accelerate drug-target exploration in S. aureus and provides critical insights into the development of base-editing tools in other microbes.

References

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Citations

Feb 11, 2019·Biotechnology and Bioengineering·Qi LiSheng Yang
Apr 19, 2019·The CRISPR Journal·Bei YangJia Chen
May 28, 2019·Human Mutation·Jinling TangTao Sun
Sep 5, 2020·Microbial Cell Factories·Zhenquan LiuDawei Zhang
Dec 25, 2019·Science China. Life Sciences·Yawei ZhaoYinhua Lu
Jul 16, 2019·Cellular and Molecular Life Sciences : CMLS·Hongming YuanHongsheng Ouyang
Jun 24, 2020·Nature Biotechnology·Andrew V AnzaloneDavid R Liu
Apr 27, 2020·Microbial Cell Factories·Yufeng LuoJianzhong Jeff Xi
Jan 15, 2021·Proceedings of the National Academy of Sciences of the United States of America·Savio D RodriguesLaurens Pauwels
Jul 6, 2021·Frontiers in Microbiology·Longxiang WangDeqiang Duanmu
Jun 22, 2021·Access Microbiology·Ian R Monk, Timothy P Stinear
Jul 2, 2020·ACS Synthetic Biology·Peng-Fei XiaBastian Molitor

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Methods Mentioned

BETA
deamination
PCR

Software Mentioned

pnCasSA
pCasSA
Integrative Genomics Viewer ( IGV )
Circos

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