用于对微管动态结构实时定量分析的荧光探针

微管是一种高度动态变化的细胞骨架成分，微管的动态不稳定性为细胞的各种活动提供了可塑性。微管动态结构与功能的异常与许多人类重大疾病密切相关。但是，目前基于小分子化合物的微管探针工具，无法精确地实时定量分析微管的动态结构和功能。本项目从当前微管探针的局限性和迫切需求出发，创新性地利用噬菌体多肽淘选和细胞穿膜肽等策略，开发能够实时动态分析微管动态结构与功能的体外单分子、活细胞和活体探针，并结合基因密码子扩展技术、蛋白共表达体系等体外和体内研究手段，突破“微管蛋白密码”对微管动态性调控研究的瓶颈，在体外单分子、活细胞和活体水平上深入研究微管动态性的调控机制，探讨微管探针作为阿尔茨海默症等疾病发生发展诊疗工具的可行性。本项目将为深入理解微管动态结构和功能的调控机制以及相关疾病的诊治提供有力工具。

Microtubules are one of the major cytoskeletal components with highly dynamics, and the dynamic property of microtubules is essential for cellular plasticity. Aberrant microtubule structures and functions are associated with many severe human diseases. However, microtubule targeting inhibitor-based probes interfere with microtubule dynamics, which challenges real-time imaging of microtubule dynamics and behaviors. Based on the current limitations and urgent needs of novel microtubule probes, this project will innovatively use strategies such as phage-displayed random peptide library panning and cell-penetrating peptides to develop in vitro single molecule, living-cell, and in vivo microtubule-targeting probes that can real-time monitor the dynamic structure and function of microtubules. Combined with in vitro and in vivo approaches, such as genetic code expansion technology and protein co-expression systems, this project will in-depth investigate the regulatory mechanisms of microtubule dynamics by the "tubulin codon" in multiple levels including in vitro single molecules, living cells and organisms. In addition, this project will explore the feasibility of microtubule probes as visual tools for the diagnosis and treatment of the development and progression of diseases such as Alzheimer’s disease. This project will help to in-depth understand the dynamic structures and functions of microtubules in physiological and pathological conditions, providing visual tools for the diagnosis and treatment of microtubule-related diseases.