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Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion
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 metho...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royal Society of Chemistry
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7069399/ https://www.ncbi.nlm.nih.gov/pubmed/32206285 http://dx.doi.org/10.1039/c9sc03784e |
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author | Zhang, Ya Zhang, Hongyuan Wang, Zhipeng Wu, Zhaowei Wang, Yu Tang, Na Xu, Xuexia Zhao, Suwen Chen, Weizhong Ji, Quanjiang |
author_facet | Zhang, Ya Zhang, Hongyuan Wang, Zhipeng Wu, Zhaowei Wang, Yu Tang, Na Xu, Xuexia Zhao, Suwen Chen, Weizhong Ji, Quanjiang |
author_sort | Zhang, Ya |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7069399 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-70693992020-03-23 Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion Zhang, Ya Zhang, Hongyuan Wang, Zhipeng Wu, Zhaowei Wang, Yu Tang, Na Xu, Xuexia Zhao, Suwen Chen, Weizhong Ji, Quanjiang Chem Sci Chemistry 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. Royal Society of Chemistry 2020-01-06 /pmc/articles/PMC7069399/ /pubmed/32206285 http://dx.doi.org/10.1039/c9sc03784e Text en This journal is © The Royal Society of Chemistry 2020 http://creativecommons.org/licenses/by-nc/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported Licence (CC BY-NC 3.0) |
spellingShingle | Chemistry Zhang, Ya Zhang, Hongyuan Wang, Zhipeng Wu, Zhaowei Wang, Yu Tang, Na Xu, Xuexia Zhao, Suwen Chen, Weizhong Ji, Quanjiang Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion |
title | Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion
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title_full | Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion
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title_fullStr | Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion
|
title_full_unstemmed | Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion
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title_short | Programmable adenine deamination in bacteria using a Cas9–adenine-deaminase fusion
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title_sort | programmable adenine deamination in bacteria using a cas9–adenine-deaminase fusion |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7069399/ https://www.ncbi.nlm.nih.gov/pubmed/32206285 http://dx.doi.org/10.1039/c9sc03784e |
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