细胞膜定位功能核酸探针的构建及其在细胞外微环境荧光成像应用的研究

Real-time monitoring of important compounds in the cellular microenvironment would provide valuable information for studying the mechanisms underlying various biological processes, such as cell metabolism, proliferation, apoptosis and signaling. Despite the significance of these extracellular local surroundings, unfortunately, their investigation has lagged far behind due to the lack of a desirable analysis technique. In this project, we propose a simple, efficient, biocompatible and universal strategy for one-step self-construction of cell-surface sensors using diacyllipid-DNA conjugates as the building elements. Based on the hydrophobic interaction between the diacyllipid and the cellular phospholipid layer, various fluorescent oligonucleotide probes with good sensing performance can be engineered onto the cell surface and used for signaling specific analytes in the cellular microenvironment with high spatiotemporal resolution. This project is expected to develop a series of cell surface-based biosensing platforms for real-time monitoring of the cellular microenvironment, providing potentially useful tools for versatile applications in cell biology, disease diagnosis, personalized therapy and regenerative medicine.

细胞外微环境中重要信息物质的实时监测对于细胞生理活动（如细胞代谢、繁殖、分化、凋亡及细胞间通讯等）机理的研究有着重要的意义。针对目前缺乏简便有效的细胞外微环境检测分析方法的现状，本项目拟以二酰基脂质为细胞膜定位单元，结合核酸适配体、脱氧核酶及G-四联体等功能核酸探针优异的分子识别性能，设计和构建多种基于细胞膜表面的荧光生物传感体系，以获取细胞外微环境中特定信息物质的实时分布信息。本项目的完成将为细胞外微环境的研究提供一个高时空分辨的荧光传感平台。同时，对细胞外微环境中重要信息物质的实时监测将为疾病的早期诊断、个性化治疗及疾病机理等领域研究提供有效的分析手段和技术支撑。

为了解决实时监测细胞外微环境中信息物质所面临的诸多问题，本项目利用细胞膜磷脂双分子层的疏水特性，结合核酸适配体、G-四联体等功能核酸探针优异的分子识别性能，设计和构建了多种基于细胞膜表面的荧光生物传感体系，实现了对细胞外微环境中特定信息物质的实时检测，为疾病的早期诊断、个性化治疗及疾病机理等领域研究提供有效的分析手段和技术支撑。本项目开展了系列的创新性工作，圆满完成了项目计划的任务，取得了丰硕的研究成果。在本项目支持下发表SCI 源刊论文7篇，其中影响因子大于5的论文6篇，包括Cell press旗下的Chem 1篇，Small 1篇，Anal. Chem. 2篇， Chem. Commun.2篇。在人才培养方面，项目负责人入选2016-2018年度中国科协“青年人才托举工程”，2018年入选“湖湘青年英才”计划。由于项目的顺利进展和初步成果，2名项目参与者获得国家优秀博士奖学金；1名项目参与者获得了国家优秀硕士奖学金；2名项目参与者获得国家留学基金委的资助，赴美国进行交流学习。