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Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion

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Abstract

Precise genetic manipulation is vital to studying bacterial physiology, but is difficult to achieve in some bacterial species due to the weak intrinsic homologous recombination (HR) capacity and lack of a compatible exogenous HR system. Here we report the establishment of a rapid and efficient method for directly converting adenine to guanine in bacterial genomes using the fusion of an adenine deaminase and a Cas9 nickase. The method achieves the conversion of adenine to guanine via an enzymatic deamination reaction and a subsequent DNA replication process rather than HR, which is utilized in conventional bacterial genetic manipulation methods, thereby substantially simplifying the genome editing process. A systematic screening targeting the possibly editable adenine sites of cntBC, the importer of the staphylopine/metal complex in Staphylococcus aureus, pinpoints key residues for metal importation, demonstrating that application of the system would greatly facilitate the genomic engineering of bacteria.

Graphical abstract: Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion

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Article information


Submitted
31 Jul 2019
Accepted
06 Jan 2020
First published
06 Jan 2020

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2020, Advance Article
Article type
Edge Article

Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion

Y. Zhang, H. Zhang, Z. Wang, Z. Wu, Y. Wang, N. Tang, X. Xu, S. Zhao, W. Chen and Q. Ji, Chem. Sci., 2020, Advance Article , DOI: 10.1039/C9SC03784E

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