Related Experiment Video
Updated: Mar 20, 2026
09:20
Reliably Engineering and Controlling Stable Optogenetic Gene Circuits in Mammalian Cells
Published on: July 6, 2021
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Single-cell characterization of bacterial optogenetic Cre recombinases.
Hellen Huang1,2, Fereshteh Jafarbeglou3,2, Mary J Dunlop1,3,2
1Molecular Biology, Cell Biology & Biochemistry Graduate Program, Boston University, Boston, MA, USA.
Iscience
|March 19, 2026
Summary
Bacterial optogenetic tools show promise for single-cell studies, but performance varies. This research compared three optogenetic Cre recombinase variants, revealing significant cell-to-cell differences in activity and timing.
Area of Science:
- Microbiology
- Synthetic Biology
- Genetics
Background:
- Optogenetic tools offer precise control over gene expression in microbial systems.
- Current bacterial optogenetic systems are often evaluated at the population level, overlooking single-cell variability.
Purpose of the Study:
- To compare the population-level and single-cell performance of three optogenetic Cre recombinase variants (OptoCre-REDMAP, OptoCre-Vvd, and PA-Cre).
- To assess the impact of reporter systems on optogenetic tool performance.
- To understand the heterogeneity of optogenetic tool activity at the single-cell level.
Main Methods:
- Utilized reporter systems measuring fluorescence or antibiotic resistance changes.
- Quantified recombination efficiency, expression variability, and activation dynamics.
- Performed single-cell analysis to evaluate heterogeneity.
Main Results:
- Optogenetic recombinase performance was found to be dependent on the chosen reporter system.
- Single-cell analysis demonstrated significant heterogeneity in recombination efficiency and activation timing.
- Substantial cell-to-cell variation was observed in the activity of optogenetic Cre recombinase variants.
Conclusions:
- Reporter choice is critical for evaluating optogenetic tool performance.
- Significant optimization is needed to enhance the single-cell capabilities of bacterial optogenetic tools.
- Findings provide criteria for selecting optogenetic recombinases for precise single-cell applications.
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