Base editor-mediated large-scale screening of functional mutations in bacteria for industrial phenotypes

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外文摘要：Base editing,the targeted introduction of point mutations into cellular DNA,holds promise for improving genome-scale functional genome screening to single-nucleotide resolution.Current efforts in prokaryotes,however,remain confined to loss-of-function screens using the premature stop codons-mediated gene inactivation library,which falls far short of fully releasing the potential of base editors.Here,we developed a base editor-mediated functional single nucleotide variant screening pipeline in Escherichia coli.We constructed a library with 31,123 sgRNAs targeting 462 stress response-related genes in E.coli,and screened for adaptive mutations under isobutanol and furfural selective conditions.Guided by the screening results,we successfully identified several known and novel functional mutations.Our pipeline might be expanded to the optimization of other phenotypes or the strain engineering in other microorganisms.

外文关键词：

base editorhigh-throughputlarge-scale screeningEscherichia colifunctional mutation

作者：

Yaomeng Yuan、Xihao Liao、Shuang Li、Xin-Hui Xing、Chong Zhang

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作者单位：

MOE Key Laboratory for Industrial Biocatalysis,Institute of Biochemical Engineering,Department of Chemical Engineering,Tsinghua University,Beijing 100084,China

Institute of Biopharmaceutical and Health Engineering,Tsinghua Shenzhen International Graduate School,Shenzhen 518055,China

Institute of Biomedical Health Technology and Engineering,Shenzhen Bay Laboratory,Shenzhen 440300,China

Center for Synthetic and Systems Biology,Tsinghua University,Beijing 100084,China

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基金：

National Key Research and Development Program of ChinaNational Natural Science Foundation of China

项目编号：

2018YFA0901500U2032210

出版年：

2024

DOI：

10.1007/s11427-023-2468-y

中国科学:生命科学(英文版)

中国科学院

中国科学:生命科学(英文版)

CSTPCD

影响因子：0.806

ISSN：1674-7305

年,卷(期)：2024.67(5)