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L-SCRaMbLE as a tool for light-controlled Cre-mediated recombination in yeast

Author

Listed:
  • Lena Hochrein

    (University of Potsdam
    University of Potsdam)

  • Leslie A. Mitchell

    (New York University Langone School of Medicine)

  • Karina Schulz

    (University of Potsdam
    Max Planck Institute of Molecular Plant Physiology)

  • Katrin Messerschmidt

    (University of Potsdam)

  • Bernd Mueller-Roeber

    (University of Potsdam
    Max Planck Institute of Molecular Plant Physiology)

Abstract

The synthetic yeast genome constructed by the International Synthetic Yeast Sc2.0 consortium adds thousands of loxPsym recombination sites to all 16 redesigned chromosomes, allowing the shuffling of Sc2.0 chromosome parts by the Cre-loxP recombination system thereby enabling genome evolution experiments. Here, we present L-SCRaMbLE, a light-controlled Cre recombinase for use in the yeast Saccharomyces cerevisiae. L-SCRaMbLE allows tight regulation of recombinase activity with up to 179-fold induction upon exposure to red light. The extent of recombination depends on induction time and concentration of the chromophore phycocyanobilin (PCB), which can be easily adjusted. The tool presented here provides improved recombination control over the previously reported estradiol-dependent SCRaMbLE induction system, mediating a larger variety of possible recombination events in SCRaMbLE-ing a reporter plasmid. Thereby, L-SCRaMbLE boosts the potential for further customization and provides a facile application for use in the S. cerevisiae genome re-engineering project Sc2.0 or in other recombination-based systems.

Suggested Citation

  • Lena Hochrein & Leslie A. Mitchell & Karina Schulz & Katrin Messerschmidt & Bernd Mueller-Roeber, 2018. "L-SCRaMbLE as a tool for light-controlled Cre-mediated recombination in yeast," Nature Communications, Nature, vol. 9(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-017-02208-6
    DOI: 10.1038/s41467-017-02208-6
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