An optogenetic toolkit for light-inducible antibiotic resistance

Michael B. Sheets, Nathan Tague and Mary J. Dunlop ()
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Michael B. Sheets: Boston University
Nathan Tague: Boston University
Mary J. Dunlop: Boston University

Nature Communications, 2023, vol. 14, issue 1, 1-13

Abstract: Abstract Antibiotics are a key control mechanism for synthetic biology and microbiology. Resistance genes are used to select desired cells and regulate bacterial populations, however their use to-date has been largely static. Precise spatiotemporal control of antibiotic resistance could enable a wide variety of applications that require dynamic control of susceptibility and survival. Here, we use light-inducible Cre recombinase to activate expression of drug resistance genes in Escherichia coli. We demonstrate light-activated resistance to four antibiotics: carbenicillin, kanamycin, chloramphenicol, and tetracycline. Cells exposed to blue light survive in the presence of lethal antibiotic concentrations, while those kept in the dark do not. To optimize resistance induction, we vary promoter, ribosome binding site, and enzyme variant strength using chromosome and plasmid-based constructs. We then link inducible resistance to expression of a heterologous fatty acid enzyme to increase production of octanoic acid. These optogenetic resistance tools pave the way for spatiotemporal control of cell survival.

Date: 2023
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DOI: 10.1038/s41467-023-36670-2

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