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Updated: Jun 21, 2025

Design and Implementation of an Automated Illuminating, Culturing, and Sampling System for Microbial Optogenetic Applications
Published on: February 19, 2017
Protocol to probe how promoters decode TF dynamics in Saccharomyces cerevisiae by combining optogenetic control with
Kieran Sweeney1, Emily L Luffey1, Megan N McClean2
1Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53705, USA.
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Gene promoters filter transcription factor (TF) localization dynamics and changes in the DNA binding affinity of TFs. Here, we present a protocol to probe how promoters decode TF dynamics in Saccharomyces cerevisiae by combining optogenetic control with microscopy. We describe steps for preparing and characterizing a light delivery platform and light-controlled TF mutants. We then detail procedures for subjecting the TFs to light doses that generate defined patterns of localization while measuring fluorescent reporter gene activation via live-cell microscopy. For complete details on the use and execution of this protocol, please refer to Sweeney and McClean.1.

