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

结构生物学提供了蛋白质与相互作用伙伴的原子视图，并且当蛋白质样品从其天然状态分离时，允许映射生理相关的相互作用物。新兴的CRISPR敲入技术能够对内源性蛋白质进行位点特异性标记，以促进从天然细胞中纯化，从而实现多蛋白质复合物的高通量结构生物学领域。该项目旨在开发一种工作流程，将CRISPR内源性标记与cryo-EM结构生物学结合起来，研究与遗传缺陷相关的超分子复合物，其中高分辨率蛋白质相互作用图谱将深入了解知之甚少的疾病机制，并为潜在的治疗靶点提供信息。

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.