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Updated: Jul 27, 2025
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Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation
Published on: October 4, 2024
637
Comprehensive screening of a light-inducible split Cre recombinase with domain insertion profiling
Biorxiv : the Preprint Server for Biology
|June 9, 2023
Summary
Researchers developed a rapid method to create split proteins for controllable function using pooled libraries and sequencing. This approach streamlines the engineering of stimulus-responsive proteins for post-translational control.
Area of Science:
- Biochemistry
- Molecular Biology
- Synthetic Biology
Background:
- Controlling protein function post-translationally is crucial for biological research and therapeutics.
- Existing methods for engineering stimulus-responsive split proteins are labor-intensive and require specialized expertise.
- Optogenetic and chemical dimers enable inducible control but require efficient split protein generation.
Approach:
- Developed a pooled library approach for parallel screening of numerous split protein constructs.
- Utilized sequencing for high-throughput readout of split protein library performance.
- Applied the method to Cre recombinase with optogenetic dimers to map split sites comprehensively.
Key Points:
- The pooled library method significantly accelerates the generation and screening of split protein variants.
- Comprehensive data on split sites for Cre recombinase with optogenetic dimers were obtained.
- A Bayesian computational approach was developed to enhance the accuracy of split protein behavior prediction.
Conclusions:
- This streamlined method facilitates the creation of inducible split proteins for precise post-translational control.
- The approach reduces the need for extensive protein engineering expertise and laborious screening.
- Enables broader application of stimulus-responsive protein control in various biological systems.
