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

CRISPR介导的内源蛋白标记用于相互作用的结构图谱

• 批准号: 2877261
• 金额: --
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2877261
• 关键词: CRISPR; mediated; tagging; endogenous; proteins

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.

结构生物学提供了蛋白质与相互作用伙伴的原子视图，当蛋白质样品从其天然状态中分离出来时，可以绘制生理相关相互作用物的图谱。 CRISPR 敲入这一新兴技术能够对内源蛋白进行位点特异性标记，从而促进从天然细胞中的纯化，迈向多蛋白复合物的高通量结构生物学领域。该项目旨在开发一种将 CRISPR 内源标记与冷冻电镜结构生物学相结合的工作流程，以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人们知之甚少的疾病机制，并为潜在的治疗靶点提供信息。