Chemically Inducible Dimerization and Chemically Disruptable Systems for Spatial and Temporal Control of Cellular Processes

Abstract

Spatial proximity–defined as the physical distance between two biomolecules–plays an important role in controlling and regulating many cellular functions. Engineered cellular systems have been developed where the physical distance between two interacting pairs can be controlled with exogenous inputs. By constraining proteins of interest to a restricted space, the relative concentration of one protein as seen by another is increased, which can lead to increased reaction rates or binding events. Recently, our group developed a new CID system called Pleiotropic Response Outputs from a Chemically Inducible Single Receiver (PROCISiR), which allows the rapid co-localization of two proteins with clinically-approved drugs. While PROCISiR is capable of some level of reversibility, the rates observed are not useful for some applications. The first chapter of this thesis describes efforts to achieve a more rapidly reversible PROCISiR system by incorporating drug-resistant mutants of the NS3a protease. The second chapter of this thesis describes efforts to increase the affinity of a previously-developed chemically-disrupted proximity system. Together, these efforts help expand the chemical genetic toolbox for rapidly controlling cellular proximity.