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bioRxiv · 10.1101/2023.04.07.536078

Lustro: High-throughput optogeneticexperiments enabled by automation anda yeast optogenetic toolkit

McClean, M. N.·Harmer, Z. P.
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Abstract

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 Saccharomyces cerevisiae. 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 high-throughput characterization of optogenetic systems across a range of biological systems and applications.

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McClean, M. N., Harmer, Z. P.. 2023-04-07. Lustro: High-throughput optogeneticexperiments enabled by automation anda yeast optogenetic toolkit. https://doi.org/10.1101/2023.04.07.536078

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