Grant: Catalyst Grant: Invention and High-Risk, High-Benefit Research

Very few of the molecular tools employed in biochemical and health research have had greater impact than intrinsically fluorescent proteins from marine organisms. The availability of these colorful proteins allows biologists to use well-established molecular biology techniques to introduce a fluorescent label into cells or tissues in a rational and chemically precise manner. With the label in place, the localization of specific proteins, that would normally be indistinguishable to our eyes, is revealed by a bright fluorescence glow that can be readily visualized using specialized microscopes. While labeling applications are the primary use of fluorescent proteins, these robust and versatile tools can also be used for the invention and development of biosensors that can actively report on biochemical activities occurring within the cell. For example, biosensors based on fluorescent proteins can report on the concentration of important ions or molecules or on the activity of particular enzymes. Our research group has recently used protein engineering to develop a set of new fluorescent proteins that enable us to image two biosensors inside of a single cell. This breakthrough has opened the door to a wide variety of new research applications since we can now monitor two biochemical activities simultaneously to investigate causality and kinetics in individual cells. We now propose to construct a series of new biosensors that will allow us to image intracellular calcium ion concentrations in live cells with an unprecedented level of spatiotemporal precision. This work will produce a toolbox of dual biosensors that will be accessible to researchers across Canada and around the globe. From this toolbox, a researcher will be able to choose a binary combination of biosensors and investigate their favorite biological question from a new perspective, ultimately leading to the medical breakthroughs of tomorrow.