Protocol for identifying glycosylated RNAs using solid-phase chemical conjugation and glycosidase release
RNA glycosylation plays a critical role in cellular signaling. Here, we present a protocol for enriching glycoRNAs from cultured cells and clinical tissues using a solid-phase chemoenzymatic technique. We describe steps for employing galactose oxidase (GAO) to selectively oxidize terminal sugar residues, enabling covalent capture on hydrazide resin, followed by enzymatic release using PNGase F. This scalable and robust assay efficiently enriches N-linked glycoRNAs, facilitating the systematic characterization of disease-specific RNA glycosylation patterns.
Authors
- Junhong Jiang (ORCID: https://orcid.org/0000-0002-2744-6710)
- Shuang Yang (ORCID: https://orcid.org/0000-0001-7958-0594)
- Xiaotong Wang (ORCID: https://orcid.org/0000-0002-6509-7734)
- Jianbo Deng
- Yu Gao
- Shengye Wen
- Zeyang Zhou
- Jiajia Li
- Xiaodong Yang
- Xinyu Miao
Institutions
- Shantou University (CN)
- Soochow University (CN)
- Shantou University Medical College (CN)
- Second Affiliated Hospital of Soochow University (CN)
- The Fourth Affiliated Hospital of Soochow University (Suzhou Dushu Lake Hospital)
Publication Details
- Journal
- STAR Protocols
- Published
- 2026-08-20
- DOI
- https://doi.org/10.1016/j.xpro.2026.104795
- Primary Topic
- RNA Interference and Gene Delivery
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Shantou University Medical College
- Suzhou University