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Title: “Conditionally Stable Proteins for Single-Molecule RNA Imaging and Protein Interaction Control In Live Cells”

Advisory Committee: Michael Economo, PhD – BME (Chair) John Ngo, PhD – BME (Advisor) Alex Green, PhD – BME Zeba Wunderlich, PhD – Biology

Abstract: Many multi-protein complexes in the cell, such as the ribosome, consist of proteins that degrade if they do not form a complex, so excess protein is eliminated if more is produced than needed for complex assembly. This work will develop methods for artificial design of proteins that degrade unless they form a specific complex, which will be applied to produce a conditionally stable RNA binding protein for live-cell single-molecule mRNA imaging with low background signal. Additionally, targeted protein degraders will be made to remove excess protein subunits during the formation of multi-protein complexes, replicating cellular function. Degradation-sensitive transcriptional and translational feedback mechanisms will be built to regulate expression of conditionally stable proteins for consistent behavior in multiple cell types. These efforts will produce a toolbox of degradation based methods for precise observation and manipulation of protein interactions.