Explore RNA Glycosylation with GlycoRNA-seq, Mass Spectrometry, and Imaging

Uncover the hidden glycan code of RNA with CD Genomics' RNA glycosylation research comprehensive service.

Our integrated platform combines GlycoRNA-seq sequencing, GlycoRNA mass spectrometry, and GlycoRNA gel electrophoresis and blot imaging, providing a comprehensive workflow for studying RNA-linked glycan modifications.

We empower academic laboratories, CROs, and pharmaceutical R&D teams to explore how RNA glycosylation influences immune regulation, RNA stability, and intercellular communication. By combining advanced chemistry and high-throughput analytics, CD Genomics enables you to transition from glycan discovery to biological insight with precision and reproducibility.

Why choose our platform:

  • Dual-labeling flexibility: Ac₄ManNAz for metabolic labeling or rPAL for post-extraction coupling
  • High-sensitivity quantification of N- and O-linked glycans from minimal input RNA
  • Integrated analysis pipeline combining sequencing, LC–MS/MS, and imaging data
  • Applicable to cells, tissues, plasma, or purified RNA samples
  • Designed for research reproducibility, QC traceability, and publication-ready results
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RNA glycosylation research platform diagram showing Ac4ManNAz and rPAL labeling workflows integrated with GlycoRNA-seq, mass spectrometry, and gel blot imaging for comprehensive RNA glycosylation analysis.
  • Ac₄ManNAz & rPAL Dual Labeling Techniques
  • RNA Glycosylation Sequencing (GlycoRNA-seq)
  • High-Resolution Mass Spectrometry of GlycoRNA
  • Gel Electrophoresis & Blot Imaging Validation
Background Labeling Strategies Service Packages Workflow Applications Advantages Sample Requirements Demo FAQ Inquiry

The Emerging Frontier of RNA Glycosylation

For decades, glycans were thought to modify only proteins and lipids. That view changed when Bertozzi's group first reported GlycoRNA in Cell (2021), identifying RNA glycosylation as a previously overlooked layer of molecular regulation.

GlycoRNAs—RNAs covalently linked to specific glycans—are now recognized as active participants in cell signaling, immune evasion, and RNA–protein interactions.

Recent discoveries have revealed that GlycoRNAs:

However, the biological functions of RNA glycosylation remain largely uncharted. Studying these low-abundance modifications requires ultra-sensitive methods and coordinated sequencing–mass spectrometry strategies.

At CD Genomics, we address this challenge by integrating GlycoRNA-seq, glycan-targeted mass spectrometry, and imaging-based validation into one cohesive workflow.

Our platform provides the technical depth needed to decode the complexity of RNA glycosylation across diverse biological systems.

Dual Labeling Strategies for GlycoRNA Detection

Understanding RNA glycosylation begins with precise and efficient labeling.

CD Genomics applies two validated chemistries—Ac₄ManNAz-based metabolic labeling and rPAL-based chemical labeling—to capture the full range of GlycoRNA molecules.

Ac₄ManNAz Metabolic Labeling

  • Introduces an azide-modified sugar (Ac₄ManNAz) into living cells.
  • Enables in-vivo incorporation of azido glycans onto newly synthesised RNA.
  • Enriches targets via biotin–streptavidin affinity for sequencing or LC–MS analysis.
  • Recommended for live-cell or dynamic metabolic studies.

rPAL Chemical Labeling

  • Suitable for purified RNA or tissue samples.
  • Uses mild oxidation to activate RNA termini, followed by alkyne–azide click chemistry.
  • Compatible with fixed or archived specimens when metabolic labeling is not feasible.
  • Provides strong signal enrichment and high reproducibility across replicates.

Both strategies support downstream GlycoRNA-seq, mass spectrometry, and gel blot imaging, ensuring consistent and cross-platform data interpretation.

Service Packages for RNA Glycosylation Analysis

Our RNA glycosylation research comprehensive service offers three complementary analytical packages—sequencing, structure mapping, and visualization—providing a complete molecular perspective from detection to validation.

Package 1 | GlycoRNA-seq (glycorna sequencing)

A next-generation sequencing workflow designed to map glycosylated RNA species, profile small RNAs, and analyse differential expression.

Analysis Workflow Overview

Analysis Type Main Steps Description / Deliverables
Standard Analysis 1. Raw read filtering & quality control
2. Genome alignment
3. ncRNA database mapping
4. sncRNA length & classification statistics
5. sncRNA quantification
6. Differential expression analysis
7. Clustering of differentially expressed sncRNAs (for biological replicates)
Provides basic transcript profiling and expression quantification of GlycoRNAs and associated sncRNAs.
Advanced Analysis 1. Sequence abundance TOP20 pie chart and TOP2 bar chart
2. Custom selection (≤50 sncRNAs) for detailed analysis
3. Target gene prediction
4. GO functional annotation
5. KEGG pathway enrichment
Offers in-depth interpretation of GlycoRNA-associated pathways, gene targets, and biological relevance.

Package 2 | GlycoRNA Mass Spectrometry Analysis

Structural and quantitative profiling of RNA-linked glycans using non-targeted LC–MS/MS or MALDI workflows.

  • Detects both N- and O-linked glycans with femtomole-level sensitivity.
  • Identifies > 60 N-glycan and ≈ 10 core2-type O-glycan structures.
  • Quantifies acp³U, galQ, manQ, and co-occurring RNA modifications.
  • Provides absolute and relative quantification with integrated spectral QC.

This package is ideal for clients seeking glycosylation-site mapping, structure elucidation, or cross-validation with GlycoRNA-seq results.

Package 3 | GlycoRNA Gel Electrophoresis and Blot Imaging

  • A visual confirmation tool for GlycoRNA presence and abundance.
  • Uses formaldehyde agarose gels to separate glycosylated RNA.
  • Transfers RNA to nylon or NC membranes for probe-based hybridisation.
  • Quantifies relative expression for cross-sample comparisons.
  • Supports initial screening or validation of sequencing and MS results.

Recommended reading:

Enhance your results with our RNA Sequencing Solutions or explore complementary Mass Spectrometry Services.

Workflow

RNA glycosylation research workflow diagram showing sample preparation, Ac4ManNAz and rPAL labeling, enrichment, GlycoRNA-seq, LC–MS/MS, gel blot imaging, and bioinformatics analysis.

Research Applications of GlycoRNA

The discovery of RNA glycosylation has expanded our view of post-transcriptional regulation.

CD Genomics' GlycoRNA-seq, GlycoRNA mass spectrometry, and gel blot imaging platforms enable in-depth exploration of this modification across diverse biological contexts.

Scientific and Translational Applications

1. RNA Structure and Stability

Study how glycan attachment affects RNA folding, secondary structure, and degradation dynamics.

2. RNA–Protein and RNA–Glycan Interactions

Map glycosylation-dependent RNA–protein binding events and identify glycan motifs influencing signal transduction.

3. Immune and Inflammatory Regulation

Characterize how GlycoRNA modulates innate immune recognition and prevents unwanted activation of TLR3/TLR7 pathways.

4. Exosomal and Secretory RNA Studies

Trace the trafficking of glycosylated RNAs in extracellular vesicles and their roles in intercellular communication.

5. Cellular Stress and Apoptotic Clearance

Analyze RNA glycosylation changes during oxidative stress or programmed cell death to reveal protective mechanisms.

6. Biomarker and Therapeutic Discovery

Detect unique GlycoRNA signatures as potential indicators of disease progression or treatment response.

Supported Research Areas

Our integrated workflow supports multi-disciplinary investigations, including:

Why Choose CD Genomics

Proven Expertise in RNA Modification Research

  • 10+ years of RNA sequencing experience across transcriptomics, epitranscriptomics, and structural RNA analysis.
  • Early adopter of GlycoRNA profiling workflows, validated on diverse biological samples.
  • Supported by a multidisciplinary team specializing in bioinformatics, chemistry, and proteomics.
  • Experienced in multi-omics integration, linking RNA glycosylation data with transcriptome and proteome layers.

Quality, Reproducibility, and Data Transparency

  • Each project includes a dedicated project manager for communication and progress tracking.
  • Internal and external QC controls ensure assay consistency and cross-run comparability.
  • Comprehensive deliverables—from raw data to analysis reports—suitable for publication or downstream validation.

Collaborative Approach

CD Genomics supports academic groups, CROs, and pharmaceutical companies in every project phase—from experimental design and sample preparation to data interpretation.

Our scientists provide consultation to help you select the most suitable labeling strategy (Ac₄ManNAz or rPAL) and optimize sequencing–mass spectrometry integration for your research goals.

Sample Requirements & Quality Control

Reliable RNA glycosylation analysis depends on strict sample integrity and controlled workflows.

CD Genomics provides clear submission guidelines to help clients obtain reproducible GlycoRNA-seq, GlycoRNA mass spectrometry, and imaging results.

Sample Requirements

Sample Type Recommended Labeling Method Minimum Amount Key Notes
Cells Ac₄ManNAz ≥ 3 × 10⁶ cells / rPAL ≥ 2 × 10⁶ cells For metabolic labeling, pre-incubate medium with 100 µM Ac₄ManNAz for 24 h.
Tissue rPAL only (≥ 25 mg RNA later preserved) ~30 µg total RNA can be extracted from 25 mg tissue.
Whole Blood rPAL recommended (3–5 mL EDTA sample) Maintain samples at 4 °C; ship on dry ice.
Purified RNA Ac₄ManNAz ≥ 50 µg / rPAL ≥ 30 µg ≥ 200 ng/µL High-purity RNA without phenol contamination.

General Handling Guidelines

Quality Control & Validation

Every project undergoes multi-layer QC to ensure accuracy and traceability:

All QC records and sample metadata are included in the final report, supporting publication and regulatory documentation.

Demo

RNA glycosylation sequencing demo results showing GlycoRNA-seq read length distribution and RNA composition pie charts for small RNA analysis.

GlycoRNA gel blot imaging demo showing wild-type and knockout RNA samples across total RNA and small RNA fractions for RNA glycosylation analysis.

FAQ

References:

  1. Flynn, R.A. et al. (2021). Small RNAs are modified with N-glycans and display glycan-dependent localization and function. Cell, 184(12):3109–3124.e22.
  2. Agard, N.J., Prescher, J.A., Bertozzi, C.R. (2004). A strain-promoted [3 + 2] azide–alkyne cycloaddition for covalent modification of biomolecules in living systems. J. Am. Chem. Soc., 126(46):15046–15047.
  3. He, M., Zhou, X. & Wang, X. Glycosylation: mechanisms, biological functions and clinical implications. Sig Transduct Target Ther 9, 194 (2024).
  4. Zhang, Y., Lu, L. & Li, X. Detection technologies for RNA modifications. Exp Mol Med 54, 1601–1616 (2022).
  5. Hu B, Ma T, Zhou D, Jiao J, Xia X. GlycoRNA, a novel RNA modification. Discov Oncol. 2025 Oct 3;16(1):1809.


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  • For research purposes only, not intended for clinical diagnosis, treatment, or individual health assessments.
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