RNAs directly regulate a vast array of cellular processes, emphasizing the need for robust approaches to fluorescently label and track RNAs in living cells. Here, we develop an RNA imaging platform using the cobalamin riboswitch as an RNA tag and a series of probes containing cobalamin as a fluorescence quencher. This highly modular ‘Riboglow’ platform leverages different colored fluorescent dyes, linkers and riboswitch RNA tags to elicit fluorescence turn-on upon binding RNA. We demonstrate the ability of two different Riboglow probes to track mRNA and small noncoding RNA in live mammalian cells. A side-by-side comparison revealed that Riboglow outperformed the dye-binding aptamer Broccoli and performed on par with the gold standard RNA imaging system, the MS2-fluorescent protein system, while featuring a much smaller RNA tag. Together, the versatility of the Riboglow platform and ability to track diverse RNAs suggest broad applicability for a variety of imaging approaches.

RNA直接调节各种各样的细胞过程，从而强调了对在活细胞中进行荧光标记和跟踪RNA的可靠方法的需求。在这里，我们开发了一个使用钴胺素核糖开关作为RNA标签以及一系列含有钴胺素作为荧光猝灭剂的探针的RNA成像平台。这个高度模块化的“ Riboglow”平台利用不同的彩色荧光染料，接头和核糖开关RNA标签，在结合RNA时引发荧光开启。我们展示了两种不同的Riboglow探针在活的哺乳动物细胞中追踪mRNA和小的非编码RNA的能力。并排比较显示，Riboglow优于染料结合适体Broccoli，与金标准RNA成像系统MS2-荧光蛋白系统表现相当，同时具有更小的RNA标签。一起，