An Expanded View of RNA Glycosylation from Metabolic Probing. GlycoRNA

The discovery of glycosylated RNA (glycoRNA) has revolutionized our understanding of the glyco- conjugate universe, revealing an unexpected association between glycans and RNA. This discovery, driven by a metabolic carbohydrate reporter, per-O-Acetylated Mannose Azide (Ac4ManNAz), under- scores the unknown potential of glycoRNA. Our study extends the discovery of ”glycoRNA” with other metabolic probes such as peracetylated N-acetylglucosamine (Ac4GlcNAz) and N-acetylgalactosamine (Ac4GalNAz) probes. Through a novel pulldown and release method, we ensured a high-confidence analysis of our sugar-azido RNA, revealing potential non-conventional sugar-RNA linkages. In-depth sequencing further revealed distinct sets of enriched transcripts for each sugar probe, emphasizing the diversity of these sugar-RNA species. Overall, these findings represents a stride towards a more comprehensive and thorough understanding of glycoRNA existence and signify the need for advanced method to unravel glycoRNA linkage structure and composition.