Lustro: High-Throughput Optogenetic Experiments Enabled by Automation and a Yeast Optogenetic Toolkit

Optogenetic systems use genetically encoded light-sensitiveproteinsto control cellular processes. This provides the potential to orthogonallycontrol cells with light; however, these systems require many design-build-testcycles to achieve a functional design and multiple illumination variablesneed to be laboriously tuned for optimal stimulation. We combine laboratoryautomation and a modular cloning scheme to enable high-throughputconstruction and characterization of optogenetic split transcriptionfactors in Saccharomyces cerevisiae. We expand the yeast optogenetic toolkit to include variants ofthe cryptochromes and enhanced Magnets, incorporate these light-sensitivedimerizers into split transcription factors, and automate illuminationand measurement of cultures in a 96-well microplate format for high-throughputcharacterization. We use this approach to rationally design and testan optimized enhanced Magnet transcription factor with improved light-sensitivegene expression. This approach is generalizable to the high-throughputcharacterization of optogenetic systems across a range of biologicalsystems and applications.