A three-colour stress biosensor reveals multimodal response in single cells and spatiotemporal dynamics of biofilms

The plethora of stress factors that can damage microbial cells has evolved sophisticated stress response mechanisms. While existing bioreporters can monitor individual responses, sensors for detecting multimodal stress responses in living microorganisms are still lacking. Orthogonally detectable red...

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Published inNPJ biofilms and microbiomes Vol. 9; no. 1; p. 57
Main Authors Zoheir, Ahmed E, Sobol, Morgan S, Meisch, Laura, Ordoñez-Rueda, Diana, Kaster, Anne-Kristin, Niemeyer, Christof M, Rabe, Kersten S
Format Journal Article
LanguageEnglish
Published London Nature Publishing Group 21.08.2023
Nature Publishing Group UK
Nature Portfolio
Subjects
Biofilms
Biosensors
Cell division
Cellular stress response
Cytotoxicity
DNA damage
E coli
Flow cytometry
Fluorescence microscopy
Genotoxicity
Herbicides
Metabolic pathways
Microfluidics
Microorganisms
Microscopy
Physiology
Propanol
Proteins
Sensors
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Summary:The plethora of stress factors that can damage microbial cells has evolved sophisticated stress response mechanisms. While existing bioreporters can monitor individual responses, sensors for detecting multimodal stress responses in living microorganisms are still lacking. Orthogonally detectable red, green, and blue fluorescent proteins combined in a single plasmid, dubbed RGB-S reporter, enable simultaneous, independent, and real-time analysis of the transcriptional response of Escherichia coli using three promoters which report physiological stress (PosmY for RpoS), genotoxicity (PsulA for SOS), and cytotoxicity (PgrpE for RpoH). The bioreporter is compatible with standard analysis and Fluorescent Activated Cell Sorting (FACS) combined with subsequent transcriptome analysis. Various stressors, including the biotechnologically relevant 2-propanol, activate one, two, or all three stress responses, which can significantly impact non-stress-related metabolic pathways. Implemented in microfluidic cultivation with confocal fluorescence microscopy imaging, the RGB-S reporter enabled spatiotemporal analysis of live biofilms revealing stratified subpopulations of bacteria with heterogeneous stress responses.
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ISSN:2055-5008
2055-5008
DOI:10.1038/s41522-023-00424-1