文献类型：期刊文献

英文题名：A genetically encoded fluorescent biosensor for sensitive detection of cellular c-di-GMP levels in Escherichia coli

作者：Li, He[1];Quan, Shu[1,2];He, Wei[1,3]

机构：[1]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, Shanghai, Peoples R China;[2]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, State Key Lab Microbial Metab, Shanghai, Peoples R China;[3]Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, Ctr Excellence Mol Cell Sci, State Key Lab Mol Biol, Shanghai, Peoples R China

年份：2025

卷号：12

外文期刊名：FRONTIERS IN CHEMISTRY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001403445700001)】；

基金：The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This work was supported by the National Key Research and Development Program of China, Grant Number: 2022YFF1102900; The National Natural Science Foundation of China (NSFC), Grant Numbers: 32222049, 32201043, and 32171269; The Natural Science Foundation of Shanghai, Grant Number: 23ZR1415300; The Shanghai Frontier Science Center of Optogenetic Techniques for Cell Metabolism, Shanghai Municipal Education Commission, Grant Number: 2021 Sci & Tech 03 28; Shanghai Municipal Science and Technology Major Project.

语种：英文

外文关键词：c-di-GMP; genetically encoded fluorescent biosensor; transcription factor; MrkH; diguanylate cyclases

摘要：Cyclic di-guanosine monophosphate (c-di-GMP) acts as a second messenger regulating bacterial behaviors including cell cycling, biofilm formation, adhesion, and virulence. Monitoring c-di-GMP levels is crucial for understanding these processes and designing inhibitors to combat biofilm-related antibiotic resistance. Here, we developed a genetically encoded biosensor, cdiGEBS, based on the transcriptional activity of the c-di-GMP-responsive transcription factor MrkH. Notably, cdiGEBS can detect both low and high cellular c-di-GMP levels, with a high fluorescence dynamic change of 23-fold. Moreover, it can detect subtle changes in c-di-GMP concentrations due to variations in the expression of c-di-GMP synthesis or degradation enzymes and can distinguish different synthesis activities among WspR mutants. These capabilities allow us to apply cdiGEBS for identifying new diguanylate cyclases and evaluating chemicals that modulate c-di-GMP levels, highlighting its potential as a high-throughput tool for screening inhibitors of c-di-GMP synthesis enzymes. Overall, cdiGEBS enhances the study of c-di-GMP-regulated functions and holds the potential for screening antimicrobials targeting c-di-GMP or its synthesis enzymes.