A High-throughput Screening Approach for the Development of FRET Biosensor
Fluorescent proteins (FPs) have transformed our ability to visualize molecular activities in live cells. New biosensors based on FPs and fluorescence resonance energy transfer (FRET) could provide the next breakthrough by providing unprecedented spatiotemporal resolution. Currently, these FRET biosensors lack sensitivity and specificity–drawbacks Professor Wang will attempt to address during his Center appointment.
His team will pursue a general, high-throughput-screening strategy based on directed evolution, yeast display, and fluorescence-activated cell sorting. The near-term goal is to optimize new biosensors for the detection of membrane-type matrix metalloproteinase 1 (MT1-MMP), which appears to play a regulator role in the degradation of extracellular matrix proteins associated with the metastasis of cancer. While MT1-MMP can be detected in a wide range of human cancers, including breast cancer, it remains unclear how its activity is regulated in live cells to control cancer development.
As the team refines successive generations of biosensors, they will characterize them in vitro and in mammalian cells to assess their sensitivity and specificity. The optimized biosensor should provide both a powerful tool for monitoring MT1-MMP activities and a convenient readout for screening more efficient and specific MT1-MMP inhibitors that can be used in cancer therapy.
In principle, the research strategy used in this project can be extended readily to develop and optimize any FRET-based genetically-encoded biosensor. Its success thus holds the potential to revolutionize our ability to perform live-cell imaging and drug discovery against intracellular targets, with a transformative impact on cell biology and medicine in general.