DNA特殊结构荧光探针的设计、合成和生物活性研究

The recognition of the special structure of DNA is very important and with great significance for the studding of the basic principle of life and disease mechanisms. It is also very important to the establishment of the new methods of disease diagnosis. The project intends to research the design and synethsize of fluorescent probe molecule, and their biological activities to the special structure of DNA. The research topics include: (1) The design and synthesis of the fluorescent probes target mismatch, bulge, hairpin structure and G-quadruplex; (2) The interaction detection of probe molecule and the special structure of DNA. Firstly, we will study the bioligcal activity of the probe with fluorescence detection. Then establish the proposed model of the interaction between probes and targets by various spectroscopic methods, such as NMR studies, computer simulation and other means, which is useful to guide the further optimize and screening of molecular probes; (3) The detection of true gene mutation sequence by the probe molecules. We will study the fluorescence response of the probe molecules in the cells which have the mismatch repair mechanism deficiency. These researchs can expand the prospects for the application of probe molecules. This project is useful to study the structure of DNA polymorphism and dynamic variation, and can promote the development of the early diagnosis of diseases which is related to DNA mutations. The accomplishment of the project also has great significance to the application of molecular fluorescence imaging in life sciences and medicine.

DNA特殊结构的识别对于研究生命活动基本规律和疾病发生机制，建立疾病诊断新方法有重要的意义。本项目拟以DNA特殊结构为靶目标，进行荧光探针分子的设计、合成和生物活性研究。主要研究内容包括（1）以错配（mismatch）、突起（bulge）、发卡结构（hairpin）和G-四链体为靶标的探针分子设计与合成；（2）探针分子与DNA特殊结构相互作用的检测，通过荧光检测评价探针分子的可行性，并拟通过各光谱学方法、核磁共振研究、计算机模拟等手段建立探针与靶标的结合模型，指导分子探针的进一步优化和筛选；（3）进行探针分子对于真实基因突变序列的检测研究，检测探针分子在错配修复机制缺失的细胞中的荧光响应，拓展探针分子的应用前景。此项目的实施，对于研究DNA的结构多态性和动态性的变化规律，建立与DNA突变相关重大疾病的早期诊断方法，推动分子荧光显像技术在生命科学、医学方面的应用有着重要的学术价值和应用前景。

本项目以DNA结构为靶标，进行荧光探针的设计、合成，用于研究DNA的结构多态性和动态变化。主要研究内容包括，以G-四链核酸和非规则核酸结构为靶标进行小分子探针的设计和合成，研究其相互作用，总结其结合规律。在项目的资助下，项目组成员以G-四链核酸和DNA甲基化修饰核酸为研究目标，设计开发了一系列特异性的荧光探针，并用于活细胞检测，取得了较好的进展。其中，设计的CPT荧光探针能够特异性识别反平行四链核酸，荧光探针TSQ1能够检测活细胞内的四链核酸结构，设计了伯胺类和羟氨类荧光分子对5-醛基胞嘧啶进行检测，苯甲醛衍生物作为荧光探针实现了选择性5-羟甲基胞嘧啶的荧光测定，并通过click反应建立快速测定5-羟甲基尿嘧啶的检测方法，除此之外，还对RNA特殊结构和其它生物大分子的小分子探针也进行了探索。本项目共发表SCI论文23篇，申请发明专利4项，已获3项授权。项目负责人翁小成获得2015年湖北省楚天学子奖励计划，培养博士研究生2名，硕士研究生2名。该项目的研究对于DNA结构多态性和动态性的变化研究有着极大的帮助作用，为与DNA结构相关的疾病研究提供了重要的技术支持。