酶标签融合蛋白的多色标记染料探针

Multicolor fluorescence imaging, calling for probes with distinguishable fluorescence signals, is an important method in biology, because it enables people to simultaneously trace and detect different targets in cells. There are large number of small-molecule dyes/probes, which are common candidates for multicolor imaging. However, the targetablity of small-molecule probes is not very satisfactory. Enzyme tag is an ideal bilogical labelling technique which integrates the diversification of small-moleucle probes and the specificity of gene-encoded protein labelling. But, the problem is that there is only very limited number of fluorescent substrates of the enzyme tags. It is not easy to obtain the matching-each-other mulitcolor substrates of different enzyme tags, which is a obstacle for the applications of enzyme tag technique. In this project, we propose a modular, efficient and versatile strategy to synthesize the fluorescent substrates of SNAP Tag, CLIP Tag or HALO Tag, and thus, prepare series of such substrates as candidates for multicolor labeling and imaging. We will also design NO-sensitive or viscosity-sensitive fluorescent substrates for different enzyme tags, and utilize them in multicolor imaging and detection of NO or viscosity in different targeted subcellular organelles or near different proteins. These studies are of novelty in biology. This project will produce some important scientific results and will promote the fusion of dyes chemistry and chemical biology.

多色成像实现细胞内多个靶标的同时追踪探测，是生物研究的重要手段，其要求不同探针的荧光信号互不干扰。小分子染料探针种类多，常用于多色成像，但其细胞内定位的靶向性和稳定性有待改善。酶标签技术结合了基因编码蛋白质标记的特异性和染料探针的多样性两方面优势，是非常理想的生物标记方法。然而，不同酶标签的荧光染料底物非常稀缺，光谱性质不容易匹配，制约了该标记技术的推广。本项目提出模块化、高效、通用性的合成策略，批量开发三种酶标签SNAP Tag、HALO Tag、CLIP Tag的特异性标记荧光底物，为多色成像提供系列的备选标记物；本项目也将设计客体（如一氧化氮）或环境因子（如粘度）敏感荧光探针修饰的多色酶标签底物，利用它们进行多个亚细胞器/蛋白质水平的多色探测成像，获得以往研究不能得到的信息，具有一定的生物学原创性。本项目将产生具有影响力的科研成果，促进染料合成化学与生物学的交叉融和。

多色成像实现细胞内多个靶标的同时追踪探测，是生物研究的重要手段，其要求不同探针的荧光信号互不干扰。小分子染料探针种类多，常用于多色成像，但其细胞内定位的靶向性和稳定性有待改善。酶标签技术结合了基因编码蛋白质标记的特异性和染料探针的多样性两方面优势，是非常理想的生物标记方法。然而，不同酶标签的荧光染料底物非常稀缺，光谱性质不容易匹配，制约了该标记技术的推广。本项目提出模块化、高效、通用性的合成策略，批量开发三种酶标签SNAP Tag、HALO Tag、CLIP Tag的特异性标记荧光底物，为多色成像提供系列的备选标记物；本项目也将设计客体（如一氧化氮）或环境因子（如粘度）敏感荧光探针修饰的多色酶标签底物，利用它们进行多个亚细胞器/蛋白质水平的多色探测成像，获得以往研究不能得到的信息，具有一定的生物学原创性。本项目将产生具有影响力的科研成果，促进染料合成化学与生物学的交叉融和。