Heparan sulfate proteoglycans facilitate the assembly of clusters of glycoRNAs and cell surface RNA-binding proteins, which negatively modulate VEGF-A signalling and angiogenesis.

Nature Structural & Molecular Biology, Published online: 06 October 2026; doi:10.1038/s41594-026-01896-3We describe a proximity-based imaging strategy for tracking glycosylated RNAs (glycoRNAs) on the surface of extracellular vesicles (EVs) and find that vesicle subtypes and cellular states are characterized by distinct glycoRNA signatures. We also demonstrate that in addition to being EV cargo, glycoRNAs contribute to intercellular communication by mediating EV uptake.

Nature Structural & Molecular Biology, Published online: 06 October 2026; doi:10.1038/s41594-026-01896-3We describe a proximity-based imaging strategy for tracking glycosylated RNAs (glycoRNAs) on the surface of extracellular vesicles (EVs) and find that vesicle subtypes and cellular states are characterized by distinct glycoRNA signatures. We also demonstrate that in addition to being EV cargo, glycoRNAs contribute to intercellular communication by mediating EV uptake.

Cell-surface glycoRNAs participate in diverse biological processes, but they were investigated as pooled populations and the protein interactome of individual glycoRNA species remain unresolved. Here we develop GlycoRNA-Map, a programmable photocatalytic proximity-labeling method that maps proteins surrounding sequence-defined endogenous glycoRNAs on living cells. A sialic-acid-binding aptamer and an RNA-hybridization probe jointly recruit a fluorophore-modified oligonucleotide through dual recognition of the glycan and RNA moieties of glycoRNAs. Light-activated biotinylation followed by quantitative mass spectrometry identified 200-418 enriched proteins for each of five glycoRNAs in A549 cells. Comparison of these proteomes revealed limited overlap among different glycoRNA sequences and across cell types, demonstrating that glycoRNA-associated protein environments depend on both RNA identity and cellular context. Transforming growth factor-{beta} receptors 1 and 2 (TGFBR1 and TGFBR2) were identified in the proximal proteomes of multiple glycoRNAs. Removal of cell-surface RNAs enhanced TGF-{beta} receptor signaling and promoted cell migration, demonstrating that glycoRNAs can functionally regulate extracellular signaling by interacting with their protein neighborhoods. GlycoRNA-Map thus provides an accessible strategy for resolving sequence-specific glycoRNA-protein interactome and discovering their roles in membrane signaling.

Nature Structural & Molecular Biology, Published online: 28 September 2026; doi:10.1038/s41594-026-01897-2The authors expand the extracellular vesicle glycosylated RNA (glycoRNA) landscape and reveal distinct glycoRNA profiles across extracellular vesicle (EV) subtypes, cell types and passage numbers. They also show that glycoRNAs are not only EV cargoes but also regulators of EV uptake, uncovering a layer of vesicle-mediated communication.

Nature Structural & Molecular Biology, Published online: 28 September 2026; doi:10.1038/s41594-026-01897-2The authors expand the extracellular vesicle glycosylated RNA (glycoRNA) landscape and reveal distinct glycoRNA profiles across extracellular vesicle (EV) subtypes, cell types and passage numbers. They also show that glycoRNAs are not only EV cargoes but also regulators of EV uptake, uncovering a layer of vesicle-mediated communication.

(a, b) Quantitative PCR analysis of GAPDH mRNA in lEVs. (a) Representative amplification curves. (b) Corresponding Ct (Cycle threshold) values. Data are presented as mean ± s.d. (n = 3 biological replicates); statistical significance was assessed using one-way ANOVA; ns, not significant. (c) Confocal microscopy images of glycoRNAs in lEVs from three colon cell lines using the EV-CAP strategy. Images shown are representative of three independent experiments with similar results. Scale bar, 5 μm. (d) Quantification of the fluorescence intensity per bead (arbitrary units, a.u.) corresponding to panel c. Data were obtained from 60 beads per group across three biological replicates and are presented as mean ± s.d. Statistical significance was assessed using one-way ANOVA; **** p < 0.0001. (e) Radar plot showing log10-transformed mean fluorescence value of per group corresponding to panel c. (f) t-distributed Stochastic Neighbor Embedding (t-SNE) analysis discriminating colon cell lines based on lEV glycoRNA fluorescence signatures. (g) Precision-recall curve (PRC) for a single lEV glycoRNA species to distinguish non-tumor colon cells (CCD-18Co) from colon cancer cells (SW480, SW620). Higher area under the curve (AUC) values indicate enhanced discriminative performance. Source data (a) Representative confocal images of glycoRNAs in lEVs from SW620 cells at the indicated...

AbstractGlycosylated RNAs (glycoRNAs) are involved in immune response, cellular function, gene regulation, and disease development. We report a liquid chromatography–mass spectrometry (LC-MS) method to analyze glycans released from glycoRNAs. The developed method includes extracting RNA from mouse liver using TRIzol, digesting proteins with proteinase K under denaturing conditions, re-extracting RNA using TRIzol, releasing N-glycans by enzymatic digestion, enriching glycans by solid-phase extraction, and analyzing glycans by LC-MS. The method was optimized to detect low-abundance N-glycans from a purchased mixture of N-glycan standards, and 65 glycans were identified. We then applied the method to analyze the glycans released from the glycoRNAs in mouse liver and identified 244 N-glycans from triplicate analyses. We also detected 40 significantly changed N-glycans in the livers of 32-week-old female and male KvB2 mice. This study establishes a simple and efficient technique for characterizing N-glycans in glycoRNAs and paves the way for future studies to understand the function of glycoRNAs.

Excessive and sustained neuroinflammation can cause neuronal damage. Wang et al. show that neuronal glycoRNAs bind Siglec-11 on immune cells, suppressing NF-κB and inflammatory cytokines and protecting neurons.

Cell-surface glycoRNAs are emerging as neuroimmune signals. In this issue of Cell Chemical Biology, Wang et al. showed in co-culture that neuronal glycoRNAs engaged microglial Siglec-11, increased SHP1 phosphorylation, suppressed NF-κB-dependent cytokine production, and improved neuronal-cell survival, establishing signaling direction, downstream mechanism, and protective function for a glycoRNA-Siglec pathway.

GlycoRNAs are a newly identified class of post‑transcriptionally modified small non‑coding RNAs that carry covalently attached N‑ and O&#

Characterization of sialic-acid-containing glycoRNAs identifies a modified base in RNA as the N-glycan attachment point.

Recent discoveries show that RNA can be modified with sialylated glycans (termed glycoRNA), thus broadening our understanding of cellular glycosylation beyond traditional proteins and lipids. However,...

Mammalian cells present RNA-binding proteins on the cell surface that form clustered domains containing glycoRNAs.

N-glycans on glycoRNAs prevent innate immune sensing of endogenous small RNAs, and the natural mechanism they use demonstrates how glycoRNAs exist on the cell surface and in the endosomal network with...

Heparan sulfate proteoglycans facilitate the assembly of clusters of glycoRNAs and cell surface RNA-binding proteins, which negatively modulate VEGF-A signalling and angiogenesis.

N-glycans on glycoRNAs prevent innate immune sensing of endogenous small RNAs, and the natural mechanism they use demonstrates how glycoRNAs exist on the cell surface and in the endosomal network without inducing autoinflammatory responses.

N-glycans on glycoRNAs prevent innate immune sensing of endogenous small RNAs, and the natural mechanism they use demonstrates how glycoRNAs exist on the cell surface and in the endosomal network…

N-glycans on glycoRNAs prevent innate immune sensing of endogenous small RNAs, and the natural mechanism they use demonstrates how glycoRNAs exist on the cell surface and in the endosomal network with...