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Light-Inducible Recombinases for Bacterial Optogenetics.

Michael B SheetsWilson W WongMary J Dunlop
Published in: ACS synthetic biology (2020)
Optogenetic tools can provide direct and programmable control of gene expression. Light-inducible recombinases, in particular, offer a powerful method for achieving precise spatiotemporal control of DNA modification. However, to-date this technology has been largely limited to eukaryotic systems. Here, we develop optogenetic recombinases for Escherichia coli that activate in response to blue light. Our approach uses a split recombinase coupled with photodimers, where blue light brings the split protein together to form a functional recombinase. We tested both Cre and Flp recombinases, Vivid and Magnet photodimers, and alternative protein split sites in our analysis. The optimal configuration, Opto-Cre-Vvd, exhibits strong blue light-responsive excision and low ambient light sensitivity. For this system we characterize the effect of light intensity and the temporal dynamics of light-induced recombination. These tools expand the microbial optogenetic toolbox, offering the potential for precise control of DNA excision with light-inducible recombinases in bacteria.
Keyphrases
  • gene expression
  • escherichia coli
  • dna damage
  • single molecule
  • air pollution
  • particulate matter
  • staphylococcus aureus
  • cystic fibrosis
  • drug delivery
  • high intensity
  • nucleic acid