Chemically Triggered Probes for High-Resolution, Multiplexed Imaging

Protein-protein interactions drive fundamental cellular processes, but the breadth and depth of such interactions remain poorly understood. This is due, in part, to a lack of reliable probes for capturing biomolecule associations in live cells and tissues. This team will develop a general strategy for imaging protein interactomes in vivo using biocompatible (bioorthogonal) probes. Such tools can selectively illuminate neighboring proteins in physiologically relevant environments.

The approach builds on a versatile bioorthogonal functional group: cyclopropenone (CpO). CpO scaffolds are inert to biological functionality, but can be selectively triggered to tag neighboring proteins, marking them for long-term visualization. CpOs can be activated “on demand,” even in cell and tissue environments. They can also be applied serially, enabling longitudinal studies in biological samples. This project will establish a suite of CpO probes for illuminating protein interactions in vivo. The probes will be engineered to emit detectable signals compatible with a wide range of imaging modalities (such as optics and electron microscopy). The molecules will be further tuned to visualize and profile multiple layers of proximal proteins. The ability to “dial in” a desired range of interaction will provide unprecedented views of biomolecule networks. Interactome labeling will be assessed using super-resolution microscopy, but the probes and protocols will be compatible with an array of detection platforms and provide a more complete picture of biomolecule functions.