Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117997, Russia.
Federal Research and Clinical Center of Physical-Chemical Medicine, Moscow 119435, Russia.
Nucleic Acids Res. 2023 Apr 11;51(6):2586-2601. doi: 10.1093/nar/gkad100.
Progress in RNA metabolism and function studies relies largely on molecular imaging systems, including those comprising a fluorogenic dye and an aptamer-based fluorescence-activating tag. G4 aptamers of the Mango family, typically combined with a duplex/hairpin scaffold, activate the fluorescence of a green light-emitting dye TO1-biotin and hold great promise for intracellular RNA tracking. Here, we report a new Mango-based imaging platform. Its key advantages are the tunability of spectral properties and applicability for visualization of small RNA molecules that require minimal tag size. The former advantage is due to an expanded (green-to-red-emitting) palette of TO1-inspired fluorogenic dyes, and the truncated duplex scaffold ensures the latter. To illustrate the applicability of the improved platform, we tagged Mycobacterium tuberculosis sncRNA with the shortened aptamer-scaffold tag. Then, we visualized it in bacteria and bacteria-infected macrophages using the new red light-emitting Mango-activated dye.
RNA 代谢和功能研究的进展在很大程度上依赖于分子成像系统,包括由荧光染料和适体基荧光激活标签组成的系统。Mango 家族的 G4 适体通常与双链体/发夹支架结合,可激活绿光发射染料 TO1-生物素的荧光,为细胞内 RNA 追踪提供了很大的应用前景。在这里,我们报告了一个新的基于 Mango 的成像平台。其主要优点是光谱特性的可调谐性和适用于需要最小标签大小的小分子 RNA 分子可视化。前者的优点是由于扩展了(绿到红发射)TO1 启发的荧光染料调色板,而截断的双链体支架确保了后者。为了说明改进平台的适用性,我们用缩短的适体-支架标签标记结核分枝杆菌 sncRNA。然后,我们使用新的红色发光 Mango 激活染料在细菌和感染细菌的巨噬细胞中对其进行可视化。