细胞内核酸及其与药物分子相互作用的核磁共振方法研究

目前核酸的结构研究主要在体外展开，而核酸通常在细胞内行使其功能，且已有研究表明细胞内的复杂环境会影响核酸的结构与功能，因此在细胞内原位开展核酸研究具有重要的生物学意义。细胞内核磁共振方法（In-cell NMR）能够提供核酸在细胞内的原位构效关系以及与药物相互作用的原子分辨信息。然而，目前该领域的发展尚处于起步阶段，十分缺乏系统的同位素标记策略研究。本项目以端粒DNA的重复序列及其突变体、核糖开关等不同核酸为研究对象，选用多种同位素标记，系统评价各标记在细胞内核酸研究中的优劣，建立起细胞内核酸的原位核磁共振方法。同时探测端粒DNA在细胞内的原位构效关系及与抗癌药物的相互作用，为抗癌药物的筛选提供一种新的原位分析方法。

The structure of nucleic acids (NAs) are mostly determined in vitro. However, NAs generally function in living cells. Previous studies show that complex cellular environment affects the structure and function of NAs. Thus, studying NAs in cells is of biological significance. In-cell NMR spectroscopy can provide atomic-level information on the structure-activity relationship of nucleic acids as well as their interaction with drugs in living cells. At present, In-cell NMR of NAs still remains a primary stage and lacks systematic investigation. Here, we select different NAs such as human telomere DNA repeat sequence and its mutant, riboswitch as models and use different isotopes labeling to systemically evaluate the advantage and disadvantage of each labeling for in-cell NA NMR. The best isotope labeling combined with NMR technique will then be applied to detect the in situ structure-activity relationship of human telomere DNA and the interactions between this DNA repeats with anticancer drugs in cells, hopefully providing a new in situ analysis method for the selection of anticancer drugs.