靶向亚细胞器的荧光分子探针

Fluorescent probes based on organic dyes have attracted remarkable attention. However, most of the probes are lack of specificity in the cellular distribution, which makes the results unreliable when sensing and imaging guest in cells. This has become one of major bottlenecks in the field of fluorescent biological analysis. To overcome this problem, in the project, we will construct two versatile rhodamine-based scaffold molecules that emit at two distinct wavelengths (540, 600nm) and have the targetablity toward subcellular lysosomes and mitochondria owing to the inducing of targeting moieties. Based on the two scaffolds, through facile chemical modifications, it is convenient to develop variety of organelles-targetable probes for different analytes. This suggests a strategy to develop a lot of probes efficiently and quickly. These probes of high applicabilty will help in discover the mechanism of active analytes's effects on subcellular organelles and thus, will be valuable tools for biological researches. The works in this project will generate important results of high impact, and will be promote the applications of fluorescent dyes in biogical and medicinal fields.

有机荧光染料作为分子探针在生命科学领域的应用取得了令人瞩目的成就。然而，绝大多数客体识别荧光探针不具备细胞内分布的靶向性，致使对细胞内客体的荧光探测和成像的可信度较低，成为荧光生物分析发展的主要瓶颈之一。为此，本项目构建两个基于罗丹明染料的母体结构，它们具有易区分的荧光发射波长（540nm，600nm）,并分别引入不同的靶向性基团，使二者各自具备特异性定位在细胞溶酶体和线粒体中的功能；这两个母体结构作为化学衍生的普适性平台，经过简捷修饰，开发出多种靶向亚细胞器的客体（一氧化氮、重金属、半胱氨酸等）识别探针。通过这项研究，能够简化靶向性荧光探针的合成，快速批量开发实用性荧光探针，应用于活细胞成像，为揭示客体对亚细胞器的调控机制提供重要方法和工具。完成此项目将产生有重要科学价值和影响力的研究结果，推进荧光染料在生物医药领域的应用。

针对传统荧光探针缺乏靶向性、荧光成像不精确的挑战，开发了一系列特异靶向溶酶体、线粒体、细胞膜等亚细胞器的荧光分子探针，实现对这些亚细胞器内局部微环境因子或客体的荧光探测成像。应用这些亚细胞器靶向探针，实现对某些疾病（如动脉硬化）早期诊断，或细胞生物学（细胞凋亡等）研究提供关键信息。并进一步基于基因编码的融合蛋白质标签技术，开发了一系列蛋白质特异性靶向的荧光分子探针，对一些亚细胞器的标志性蛋白，如细胞核组蛋白、线粒体电子转移COX8A蛋白等，实现了特异性标记，并检测蛋白附近局部区域的粘度、一氧化氮等。上述研究将推进荧光探针在细胞成像、疾病诊断、药理研究等方面的应用，促进染料化学与细胞生物学的交叉融合。