SBIR Phase I: New Fluorescent Biosensors for Signaling in Living Cells

This Small Business Innovation Research (SBIR) Phase I project will establish the feasibility of developing a genetically-encoded biosensor to monitor levels of cyclic adenosine monophosphate (cAMP), an important second messenger component of drug signaling pathways. Unlike existing FRET-based biosensors that depend upon energy transfer between two fluorescent molecules, this sensor will employ a single, circularly permuted fluorescent green protein. This green sensor can be coupled with differently colored biosensors for other second messengers to produce simultaneous readouts for multiple components of a signaling pathway that change when activated by a drug. The broader impact/commercial potential of this project is that multiplex, genetically-encoded assays reporting multiple cell signaling events would expand the depth of knowledge about signal transduction by improving information on the timing, location and pathway cross-talk in physiologically relevant tissues. These assays are homogenous, do not require multiple steps, or cell lysis. A growing trend in the pharmaceutical industry is screening in primary cell cultures, and genetically encoded assays are ideally suited for this. The technology to be developed in this proposal represents a new innovation in fluorescent live-cell assay and has strong potential to reduce the cost and improve the reliability of drug discovery to find safe and effective drugs that provide better treatment outcomes and improved human health.