high-throughput-optogenetics-experiments-yeast-using-automated

High-Throughput Optogenetics Experiments in Yeast Using the Automated Platform Lustro

Optogenetics offers precise control over cellular behavior by utilizing genetically encoded light-sensitive proteins. However, optimizing these systems to ...

University of Wisconsin-Madison

One snapshot of the peer review process for "Synthetic gene networks recapitulate dynamic signal decoding and differential gene expression" (Benzinger et al., 2022).

Evaluation of Benzinger et al.: Optogenetic circuits for dynamic signal processing.

Optogenetic systems use genetically encoded light-sensitive proteins to control cellular processes. This provides the potential to orthogonally control cells with light; however, these systems require many design-build-test cycles to achieve a functional design and multiple illumination variables need to be laboriously tuned for optimal stimulation. We combine laboratory automation and a modular cloning scheme to enable high-throughput construction and characterization of optogenetic split transcription factors in . We expand the yeast optogenetic toolkit to include variants of the cryptochromes and enhanced Magnets, incorporate these light-sensitive dimerizers into split transcription factors, and automate illumination and measurement of cultures in a 96-well microplate format for high-throughput characterization. We use this approach to rationally design and test an optimized enhanced Magnet transcription factor with improved light-sensitive gene expression. This approach is generalizable to the high-throughput characterization of optogenetic systems across a range of biological systems and applications.

Article	Year
Evaluation of Benzinger et al.: Optogenetic circuits for dynamic signal processing. Cell systems\| PubMed ID: 35588696	2022
Lustro: High-Throughput Optogenetic Experiments Enabled by Automation and a Yeast Optogenetic Toolkit. ACS synthetic biology\| PubMed ID: 37434272	2023

Zachary P. Harmer

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