Systematic High-Content Screening of Fluorescently Tagged Yeast Double Mutant Strains.

We describe a protocol for high-content screening in budding yeast that can be used to study genetic interactions from a cell biological perspective. This approach can be used to map genetic interactions by monitoring one or more subcellular fluorescent markers of interest. In this case, changes in the morphology or abundance of a subcellular compartment, pathway or bioprocess are monitored in the background of a systematic array of yeast double mutants. Alternatively, the protocol can be used to monitor proteome-wide abundance and localization changes in a double mutant of interest by screening the yeast ORF-GFP collection. The protocol can be readily adapted for high-content screening of triple mutants, other large-scale yeast collections or expanded to screening of multiple growth conditions.