基于DNA催化自组装的模块化荧光生物传感技术及其生物医学应用研究

项目名称： 基于DNA催化自组装的模块化荧光生物传感技术及其生物医学应用研究

项目编号： No.21475077

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 数理科学和化学

项目作者： 姜玮

作者单位： 山东大学

项目金额： 95万元

中文摘要： 本项目拟以实现肿瘤标志物的灵敏特异性检测和多种肿瘤标志物联合检测为目标，以动态DNA纳米技术和荧光分析技术为研究平台，以Toehold介导的链置换反应为核心反应，重点解决以下关键科学问题：（1）利用目标物诱导DNA构象转变、酶催化作用、变异基因的连接或聚合等特异性反应设计构建分子识别模块，并解决与DNA三向节模块的对接问题；（2）利用Toehold的结合活化策略设计组装能实时、同步、定量转换目标分子特异性识别事件的DNA三向节模块，在提高方法通用性的同时，解决触发链二级结构多样性带来的背景泄漏问题；（3）利用碱基错配机制优化发卡的设计，解决发卡泄露造成的背景干扰问题，构建高效的单循环和指数动态DNA催化自组装荧光信号放大模块；（4）以即插即用的方式构建超灵敏、特异性、模块化的肿瘤标志物检测系统和多种肿瘤标志物的联合检测系统。以期为癌症早期筛查、准确诊断和有效治疗提供有力的技术支持。

中文关键词： DNA催化自组装；模块化；荧光检测；生物医学应用

英文摘要： In this proposal, we intend to introduce dynamic DNA nanotechnology and fluorimetric technique as the basis and toehold-mediated strand displacement as a key reaction to focus on the following scientific problems, aiming to realize sensitive and specific detection of tumor markers and combined detection of tumor marker levels. Firstly, analyte-induced DNA conformational change, the mechanism of enzyme action, and allele-specific ligation or extension will be used to design and construct molecule recognization modules which will be utilized to dock with the three-way junction modules. Secondly, three-way junction modules will be designed and assembled on the basis of the associative toehold activation, which derive from the real-time, synchronous, and determined molecule specific recognization. The introduction of associative toehold activation will enhance the universality of the research method and reduce the background leakage originating from the misfolding of catalytic DNA. Thirdly, the mismatch mechanism will be recommended to optimize the design of the hairpin substrates, decrease the background signal owing to uncatalyzed strand displacement reactions and dynamically assemble efficient linear and exponential amplified fluorescent signal modules. Finally, the architecture of detection will be integrated via modular components, which can realize sensitive and specific detection of tumor markers and combined detection of tumor marker levels in plug-and-play mode. The proposed research is expected to provide powerful technical support for the early screening, accurate diagnosis, and effective treatment of cancers in clinical environment.

英文关键词： DNA Catalyzed Hairpin Self-assembly;Modularity;Fluorescent Detection;Biomedical Application