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Updated: Jun 13, 2025
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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.
Biorxiv : the Preprint Server for Biology
|June 12, 2025
Summary
Bacterial optogenetic tools show variable performance in single cells. This study compares three optogenetic Cre recombinase variants, revealing reporter-dependent and cell-to-cell differences in activity crucial for precise genetic control.
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, masking single-cell variability.
- Stochastic effects significantly impact gene expression dynamics in individual bacterial cells.
Purpose of the Study:
- To systematically compare the population-level and single-cell performance of three optogenetic Cre recombinase variants (OptoCre-REDMAP, OptoCre-Vvd, and PA-Cre).
- To investigate the influence of reporter systems on optogenetic recombinase activity.
- To characterize the heterogeneity in activation dynamics and recombination efficiency at the single-cell level.
Main Methods:
- Utilized reporter systems measuring fluorescence or antibiotic resistance to quantify Cre activity.
- Employed single-cell analysis to assess recombination efficiency, expression variability, and activation timing.
- Compared three distinct optogenetic Cre recombinase variants under light induction.
Main Results:
- Optogenetic recombinase performance was found to be dependent on the chosen reporter system.
- Significant heterogeneity in light-induced Cre activity was observed across individual bacterial cells.
- Substantial cell-to-cell variation in the timing of recombinase activation was detected.
Conclusions:
- Reporter choice is a critical factor in designing effective optogenetic systems for bacteria.
- Current optogenetic tools exhibit considerable variability in single-cell applications, necessitating careful selection and optimization.
- Further development is needed to enhance the reliability and precision of bacterial optogenetic tools for single-cell studies.
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