Enzymatic covalent labeling of RNA with RNA transglycosylation at guanosine (RNA-TAG).

Abstract

Technologies for the labeling, detection, and manipulation of biomolecules have drastically improved our understanding of cell biology. As the myriad of functional roles for RNA in the cell are increasingly recognized, such tools to enable further investigation of RNA are the subject of much interest. RNA-TAG is an enzymatic method for site-specific, covalent labeling of RNA. This methodology makes use of a bacterial tRNA modifying enzyme, tRNA guanine transglycosylase, to incorporate modified substrate analogs into a target RNA, resulting in highly efficient and site-specific RNA labeling. In this chapter, we introduce the underlying principles of the RNA labeling reaction, discuss various applications of RNA-TAG, and present protocols for labeling specific RNA transcripts using this system.