CRISPR-mediated tagging of endogenous proteins for structural mapping of interactions

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Structural biology offers atomic views of proteins with interacting partners, and when protein samples are isolated from their native states, allows mapping of physiologically relevant interactors. The emerging technology of CRISPR knock-in enables site-specific tagging of endogenous proteins to facilitate purification from native cells, towards the realm of high-throughput structural biology of multi-protein complexes. This project aims at developing a workflow that couples CRISPR endogenous tagging with cryo-EM structural biology, to study supramolecular complexes associated with genetic defects, where high-resolution protein protein interaction maps would yield insight into the poorly understood disease mechanism and inform potential therapeutic targets.