Transcriptome-wide coverage of glycoRNA species
GlycoRNA Analysis Service
What Is GlycoRNA?
Fig.1 GlycoRNA Trafficking & Function.1,4
GlycoRNAs are RNA molecules covalently modified with N-linked glycans, previously thought to decorate only proteins and lipids. These RNAs—predominantly small noncoding types such as Y RNAs, tRNAs, and vault RNAs—carry sialylated and fucosylated glycans, localize to the cell surface, and participate in cell–cell signaling, immune regulation, and tumor microenvironment remodeling. They are synthesized via an atypical pathway involving RNA-specific glycosylation enzymes such as GALNTs, sialyltransferases, and DTWD2. GlycoRNAs may traverse through Golgi/ER-associated vesicular pathways or leverage RNA-binding protein (RBP)–mediated shuttling, ultimately appearing on the plasma membrane.
Key Types of GlycoRNAs
Currently, the known types of glycosylated non-coding RNAs (sncRNAs) include small nuclear RNAs (snRNAs), ribosomal RNAs (rRNAs), small nucleolar RNAs (snoRNAs), transfer RNAs (tRNAs), Y-RNAs, and microRNAs (miRNAs). Notably, glycosylated messenger RNAs (mRNAs) have not been identified, indicating a unique role for these modifications in non-coding RNAs rather than coding RNAs. Below, we detail the key types of GlycoRNAs:
👉 Small Nuclear RNAs (snRNAs)
Involved in RNA splicing, glycosylation can fine-tune their interaction with spliceosomal proteins, thereby affecting splicing efficiency.
👉 Ribosomal RNAs (rRNAs)
Glycans attached to rRNAs can influence ribosome structure, stability, and the cell's response to stress.
👉 Small Nucleolar RNAs (snoRNAs)
Modifications on snoRNAs may regulate their role in rRNA maturation and ribosome biogenesis.
👉 Transfer RNAs (tRNAs)
Glycosylation impacts tRNA folding, amino acid binding, and safeguards against degradation.
👉 Y-RNAs and MicroRNAs (miRNAs)
Glycans alter their stability, interaction with protein complexes, and involvement in gene silencing or DNA replication.
Fig.2 Three
types of non-coding RNAs.2,4
GlycoRNA Analysis Service Portfolio
| Service Type | Description |
|---|---|
| GlycoRNA Profiling Service | High-throughput sequencing and novel subtype discovery |
| GlycoRNA Functional Analysis | Assessment of glycoRNA-mediated signaling, immune evasion, or translation impact |
| GlycoRNA Imaging Service | Spatial visualization of glycoRNAs using FRET imaging |
Why Choose Creative Biolabs?
Whether investigating glycoRNAs in cancer biology, autoimmunity, or infection, our team delivers tailored strategies with scientific rigor and technical precision.
Creative Biolabs integrates cutting-edge transcriptomics, click chemistry, and imaging platforms to provide:
Single-cell resolution imaging of glycoRNA spatial distribution
Validated quantification of glycosylation sites
Functional assays to decode biological consequences
Why Study GlycoRNA?
GlycoRNAs offer a unique vantage point for exploring disease-specific regulatory mechanisms. Recent studies have revealed that their surface localization and interactions with Siglec receptors, lectins, and glycan-binding proteins confer them significant roles across disease categories. Below, we categorize the relevance of glycoRNAs by pathological context:
Cancer
- Tumor Immune Evasion: GlycoRNAs bind to Siglec-5 and P-selectin on immune cells, promoting immunosuppressive signaling that enables cancer cell evasion.
- Diagnostic Biomarkers: FRET imaging has demonstrated that glycoRNA profiles can distinguish between tumor types with >90% accuracy.
- Breast Cancer Progression: Expression of U1 and Y5 glycoRNAs decreases with malignancy, suggesting utility in staging and prognosis.
Autoimmune and Inflammatory Disorders
- Neutrophil-Endothelium Interaction: Cell-surface glycoRNAs facilitate leukocyte adhesion during inflammation. RNase digestion of glycoRNAs reduces monocyte and neutrophil attachment to endothelial cells.
- Macrophage Activation: Upon LPS stimulation, some glycoRNAs increase significantly, implicating them in acute inflammatory response modulation.
Infectious Disease and Viral Immunity
- Viral infections may hijack glycoRNA pathways or alter RNA glycosylation profiles. Although less explored, glycoRNA profiling could serve as a signature for immune activation stages or host-pathogen interactions, especially given their surface accessibility.
Neurological Disorders and Developmental Biology
- Emerging evidence links glycoRNAs to lipid raft microdomains in neurons. The glycan-rich microenvironment on the neuronal membrane may influence synaptic signaling or axon guidance.
Trusted by Scientists Worldwide
How to Analyze GlycoRNA?
GlycoRNA-seq: High-Throughput Transcriptome-Wide Mapping
GlycoRNA-seq platform combines metabolic labeling, bioorthogonal chemistry, and deep sequencing to enable sensitive, transcriptome-wide detection of glycosylated RNAs. Compared to earlier methods, this approach offers higher specificity, broader transcript coverage, and robust validation.
Novel GlycoRNA Imaging: Single-Cell Spatial Visualization
A Novel GlycoRNA Imaging combines sialic acid aptamers and RNA hybridization probes. It visualizes glycoRNAs with subcellular localization, revealing:
- Colocalization with lipid rafts
- Trafficking via SNARE-mediated vesicle fusion
- Abundance variations across tumor stages
Clier-qPCR: Specificity and Quantitation
Clier-qPCR enables sensitive and specific detection of glycosylated RNAs across diverse sample types. It serves as a critical validation step for sequencing-based discovery and functional assays.
Additional Methods
- rPAL labeling
- RNA-blotting with DBCO-biotin conjugates
- HPLC-MS of released glycans
Sample Requirements & Submission
To streamline your glycoRNA project, please prepare and submit samples according to the table below. For customized protocols or uncommon sample types, contact our technical support team.
| Sample Parameter | Requirement | Notes |
|---|---|---|
| Sample Type | Cells, total RNA, tissue (frozen/fixed) | Avoid protease-based dissociation for surface glycoRNAs |
| Cell Number | ≥ 1 × 10⁷ cells | Suspension or adherent lines; growth log recommended |
| Total RNA | ≥ 5 μg; OD260/280 1.8–2.0 | TRIzol, column, or phenol-chloroform extraction |
| Tissue | ≥ 30 mg or 1 section (10 μm) | Snap-frozen or OCT-embedded; avoid paraffin |
| RNA Quality | RIN ≥ 7.0 | Required for sequencing; optional for imaging |
| Preservative-free | No RNAlater, heparin, or detergents | These inhibit click chemistry or enzymatic steps |
Insights from Ongoing Studies
Breast Cancer GlycoRNA Imaging
Using a novel glycoRNA imaging technique, researchers visualized U1, Y5, and SNORD35a glycoRNAs in:
- MCF-10A (non-tumorigenic): High glycoRNA signal
- MCF-7 (malignant): Moderate signal
- MDA-MB-231 (metastatic): Low or absent signal
The inverse relationship between glycoRNA abundance and tumor aggressiveness indicates potential for glycoRNA-based diagnostics.
GlycoRNAs in Innate Immune Cells
Upon LPS stimulation, glycoRNA levels (U1, Y5) significantly increased in macrophages. Conversely, neutrophil maturation decreased glycoRNA signal. RNase treatment of immune cells impaired endothelial adhesion, supporting a functional role in immune cell–vascular interactions.
Beyond GlycoRNA: Comprehensive Glycosylation Analysis Services
Fc/Fab glycosylation profiling, sialylation analysis
Monoclonal Antibody Glycosylation Analysis Polyclonal Antibody Glycosylation Analysis Fab Glycosylation Analysis Fc Glycosylation Analysis
Glycolipid Analysis
Glycosphingolipid, glycoglycerolipids and more types of glycolipids characterization
Glycosphingolipids Analysis Glycoglycerolipids Analysis Lipopolysaccharide Analysis Glycosylphosphatidylinositol Anchor AnalysisFAQs
How can I validate that the RNA glycosylation observed in my sample is real and not an artifact?
At Creative Biolabs, we use a three-tier validation strategy to ensure specificity and eliminate false positives:
- Metabolic Labeling Controls: We always include Ac4ManNAz-unlabeled controls to determine background biotin enrichment.
- Clier-qPCR: This method calculates a corrected enrichment-to-input ratio, distinguishing true glycosylated RNAs from non-specific binding.
- Enzymatic Digestion Verification: Treatment with RNase A and PNGase F is used to confirm that both the RNA and glycan components are essential for the signal. We've demonstrated that RNase A abrogates the RNA signal, while glycosidases like PNGase F eliminate the glycan-dependent component of the glycoRNA signal.
This combination gives us high confidence in the biological authenticity of the glycoRNAs detected.
What types of RNA are typically glycosylated, and can I identify novel glycoRNA subtypes in my own cell model?
Yes. Our glycoRNA-seq pipeline allows transcriptome-wide discovery of both annotated and novel glycoRNA species. Common glycosylated RNA classes include:
- tRNAs (notably tRNA-Ser, tRNA-Thr, tRNA-Val, and tRNA-Lys)
- Vault RNAs (e.g., vtRNA2-1)
- Y RNAs and snoRNAs
- Unannotated lncRNAs
Can glycoRNA profiling support disease biomarker discovery or clinical diagnostics?
Yes. In fact, our FRET-based glycoRNA profiling has already demonstrated potential for cancer diagnostics. With our help, you can define signature glycoRNA panels for biomarker development in oncology, immunology, or inflammation research.
What Our Clients Say
References:
- Parsons, James V. "Exploring Foundational Experiments in GlycoRNA." (2024). https://www.preprints.org/manuscript/202411.0096/v1
- Good, Deborah J. "Non-coding RNAs in human health and diseases." Genes 14.7 (2023): 1429. https://doi.org/10.3390/genes14071429
- Zheng, Linlin, et al. "The glycosylation of immune checkpoints and their applications in oncology." Pharmaceuticals 15.12 (2022): 1451. https://doi.org/10.3390/ph15121451
- Distributed under Open Access license CC BY 4.0 , without modification.
Featured Products
For Research Use Only. Not For Clinical Use.
Related Services:
Online Inquiry