Chemical Biology Tools for Decoding the Cell Surface GlycoRNA

ABSTRACT The discovery of glycosylated RNAs (glycoRNAs) on the outer surface of mammalian cells, represents a spatial awakening in RNA biology. This identification redefines the spatial boundaries of RNA biology, possibly extending its role beyond the nucleus and cytoplasm to the extracellular membrane interface. This unexpected localization and the presence of glycosylation modifications challenge longstanding conventional paradigms and raise new questions about the biosynthetic pathways, structural diversity, and signaling functions of glycoRNAs in the intercellular environment. Chemical biology tools are central to their discovery, further identification, and future exploration, enabling enrichment, imaging, and functional analysis of glycoRNAs in both physiological and pathological contexts. In this review, we highlight how these technologies have driven the discovery of glycoRNAs and revealed new principles of RNA localization and membrane‐related functions. We further discuss several challenges and future directions in decoding the RNA‐centric membrane biology.