Lectin-based detection and expression profiling of native glycoRNAs

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.