Analytical methods for glycoRNA detection: a systematic review

Glycosylated RNAs (glycoRNAs) are a groundbreaking discovery in glycobiology, extending the diversity of known glycoconjugates beyond proteins and lipids. Accumulating evidence indicates their pivotal roles in cellular communication, immune regulation, and disease pathogenesis, highlighting their promise as novel biomarkers and therapeutic targets. Robust and sensitive detection methods are crucial for deciphering glycoRNA glycosylation patterns and elucidating their physiological and pathological functions. Despite the development of diverse analytical techniques, a systematic evaluation of glycoRNA detection strategies remains lacking. To address this gap, in this review, we comprehensively summarize recent methodological advances, categorizing the published approaches based on their glycan-targeting recognition mechanisms and critically assessing their principles, applications, strengths, and limitations. We also sketch the key challenges and future development directions. This review aims to provide a timely and informative guide for researchers in this rapidly evolving area.