PT  - JOURNAL ARTICLE
AU  - Ahmed E. Zoheir
AU  - Morgan S. Sobol
AU  - Diana Ordoñez-Rueda
AU  - Anne-Kristin Kaster
AU  - Christof M. Niemeyer
AU  - Kersten S. Rabe
TI  - A genetically-encoded three-colour stress biosensor reveals multimodal response at single cell level and spatiotemporal dynamics of biofilms
AID  - 10.1101/2022.09.23.509207
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.09.23.509207
4099  - http://biorxiv.org/content/early/2022/09/24/2022.09.23.509207.short
4100  - http://biorxiv.org/content/early/2022/09/24/2022.09.23.509207.full
AB  - The plethora of chemical, physical, and biological factors that can damage microbial cells has triggered the evolution of sophisticated stress response (SR) mechanisms. While individual SR pathways have been monitored with genetically encoded reporters, sensor concepts for the detection of multimodal effects of stressing conditions in living microorganisms are still lacking. Orthogonally detectable red, green, and blue fluorescent proteins combined in a single vector system, dubbed RGB-S reporter, enable the simultaneous, independent and real-time analysis of the stress response in Escherichia coli to physiological stress, genotoxicity, and cytotoxicity. The sensor system can be read out via conventional fluorescence microscopy or microtiter plate analysis and can also be combined with Fluorescent Activated Cell Sorting (FACS) and subsequent transcriptome analysis. Various stressors, such as the biotechnologically relevant 2-propanol, lead to the activation of one, two or all three SRs, which can have a significant impact on non-stress-related metabolic pathways. Implemented in microfluidic cultivation with confocal fluorescence microscopy imaging, the technology enabled spatiotemporal analysis of live biofilms to discover stratified subpopulations of bacteria with heterogeneous stress responses.Competing Interest StatementThe authors have declared no competing interest.