TRD 2: Molecular Imaging Tools for Visualizing and Controlling Cellular Responses to Force

The Hahn laboratory focuses on the development of new approaches to quantify the spatio-temporal dynamics of signaling in living cells, and to engineer genetically encoded proteins so that their specific activities can be manipulated with second and micron resolution in vivo. As a new component in CISMM, TRD 2 provides collaborators with the ability to visualize and manipulate signaling molecules in cells as they are subjected to controlled forces. We provide already-developed biosensors and photo-responsive signaling proteins, and apply well-validated methods to generate new biosensors or engineer light and small molecule sensitivity into molecules specified by CISMM collaborators. Proteins of special relevance to force and mechanics include the guanine nucleotide exchange factors (GEFs), which modulate the Rho family of GTPases and related kinases.

Project 2.1. Develop fluorescent biosensors to quantify the spatio-temporal dynamics of protein activity in living cells during the application of mechanical forces, using novel designs with enhanced sensitivity and reduced physiological perturbation.

Project 2.2.  Develop broadly applicable approaches to confer light-induced activation and release of proteins in living cells, and use these to generate localized changes in protein activity at precise positions and times relative to applied forces.

Project  2.3. Develop engineered protein domains and identify domain insertion sites to confer light-induced inhibition and rapamycin-induced activation of kinases and GEFs without sequestration.