Posttranscriptional chemical labeling of RNA by using bioorthogonal chemistry

•Posttranscriptional chemical labeling is emerging as a new method to profile RNA.•RNA labeling methods using bioorthogonal reactions are described in this review.•Methods to introduce bioorthogonal reactive groups into RNA are described first.•Utility of these techniques in profiling RNA in vitro and live cells is discussed.

Recent developments in RNA labeling technology have provided viable tools to analyze RNA synthesis, processing and function in cell-free and cellular environments. Notably, emerging methodologies based on posttranscriptional chemical labeling by using bioorthogonal chemistry have enabled the visualization and profiling of exogenous and endogenous RNA transcripts. In this review, we first give an overview of different RNA labeling strategies based on chemical as well as genetically encoded systems. Subsequently, we provided a detailed discussion on methodologies that have been developed to introduce various bioorthogonal reactive groups into RNA transcripts, which are compatible for posttranscriptional functionalization. Finally, the utility of these techniques in imaging and studying the dynamics of RNA production, distribution and decay in complex cellular environment is discussed.