基于DNA仿生酶的MicroRNA检测

MicroRNA molecules have an important role in cancer diagnostic, but it is challenging to detect these molecules without the use of time-consuming and error-prone amplification methods. Here, we present a platform based on DNA Protein mimic (DNA Protmin) for the rapid and sensitive detection of microRNAs . In this project, bio-inspired by nature protein, we aim to construct a noval microRNA responded DNA protmin, in which DNA Tetrahedron is applied as a scaffold to support G-quadruplex and enhance its catalytic activity and detection sensitivity. Meanwhile , to further improve its selectivity and specificity, we construct DNA protmin cluster with tunable tetrahedral scaffolds. By optimizing their structure and configuration, the DNA protmin and its cluster are aimed to identify the target microRNA with high sensitivity, selectivity and specificity. We hope this project provide theoretical basis and experimental model for the construction of DNA protmin with high performance and supply new materials and new technologies for cancer diagnosis and pathology studies.

MicroRNA在肿瘤诊断中有着重要的应用价值，但是传统靶标扩增方法费时耗力且误差大，给microRNA的检测带来了诸多困难。本项目拟利用DNA纳米技术和G-四联体酶，构建基于DNA仿生酶的快速、高灵敏度的microRNA检测方法。在该DNA仿生酶构建中，受到天然蛋白酶的启发，我们使用DNA四面体骨架保护G-四联体酶的结构，从而提高其催化活性以及检测灵敏度。同时，我们使用结构可调的DNA四面体骨架构建具有协同效应的DNA仿生酶簇，从而进一步提高其检测选择性和特异性。本项目期望通过优化DNA仿生酶及其酶簇的结构和构象，构建对靶标microRNA具有高灵敏度、选择性和特异性的DNA仿生酶和酶簇，从而实现其对MicroRNA的快速、高效、准确的检测。本项目有望为DNA仿生酶的构建提供理论基础和实验模型，为癌症的诊疗提供新材料和新技术。

本项目以开发高活性的DNA仿生酶为目标，以框架核酸的研究为核心，设计和制备基于核酸纳米酶，并研究框架核酸与活性中心的关系。在该项目中，创建基于框架核酸的单分子限域与多分子协同动态组装策略，提高了框架核酸仿生酶的组装效率与精度，提高核酸适体的识别能力；利用框架核酸纳尺度空间界面的动态组装策略，发展了单分子定位示踪系统，并拓展其在单分子原位化学动态测量中的应用，为相关疾病的研究奠定基础。开展的工作完成了预期的研究目标，为癌症的诊疗提供新材料和新技术。