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，当蛋白质样本与其天然状态分离时，可以绘制出生理上相关的相互作用因子图。新出现的CRISPR基因敲入技术使内源蛋白质的位点特异性标记能够促进从天然细胞中提纯，走向多蛋白质复合体的高通量结构生物学领域。该项目旨在开发一种将CRISPR内源标记与低温EM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体，其中高分辨率蛋白质相互作用图将深入了解鲜为人知的疾病机制，并为潜在的治疗靶点提供信息。