Abstract
Current methods for detecting glycoRNAs include metabolic labeling in living cells or animals and RNA-optimized periodate oxidation and aldehyde labeling (rPAL), each of which offers distinct advantages and limitations. Here, we report a relatively simple and rapid approach for detecting native glycoRNAs using direct lectin hybridization. This method involves several straightforward steps, including total RNA isolation, northern blotting, and lectin hybridization. Its advantages include high sensitivity, procedural simplicity, and broad applicability. Using this approach, we profiled glycoRNA expressions in RNA samples derived from human and murine tissues and cell lines and compared the results with those obtained using two established detection methods. We also examined differences in glycoRNA expression under physiological and pathological conditions. Notably, we report for the first time the detection of free glycoRNAs in various human biofluids, including plasma, urine, and amniotic fluid. Overall, our findings demonstrate that this method is reliable and reproducible, providing an alternative tool for studying glycoRNA biology and potentially offering utility for future clinical diagnostics.
Data availability
The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We thank Drs. Tony Wang, Daping Fan, Kun Chen, Jianming Qiu, Shizhen Wang and Cuthbert Simpkins for kindly providing cell lines and/or RNA samples. This project was supported by the National Institutes of Health (R15AI138116, to M.F), UMKC Funding for Excellence (2023, to M.F), SPiRE (2024, to M.F) and UMKC SOM Bridge Fund (2025, to M.F).
Funding
This work was supported by the National Institutes of Health (R15AI138116 to M.F.), UMKC Funding for Excellence (2023 to M.F.), SPiRE (2024 to M.F.), and the UMKC School of Medicine Bridge Fund (2025 to M.F.).
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M.F. conceived, designed and supervised the overall study. Y.L., X.L. and M.F. performed most of the experiments. Y.Q. performed metabolic labeling experiments for mice. T.L. and P.N. help to perform mouse tissue expression experiments, and M.F. wrote the manuscript. M.F., H.M. and P.N. revised the manuscript.
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Li, Y., Qian, Y., Li, X. et al. Lectin-based detection and expression profiling of native glycoRNAs. Sci Rep (2026). https://doi.org/10.1038/s41598-026-40291-2
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DOI: https://doi.org/10.1038/s41598-026-40291-2
