RNA-associated glycoconjugates highlight potential ambiguities in glycoRNA analysis
A recent ground-breaking study suggested that small RNA from mammalian cells can undergo N-glycan modifications (termed glycoRNA)<sup>1</sup>. The discovery relied upon a metabolic glycan labeling strategy in combination with commonly used phase-separation-based RNA isolation. Following the reported procedure, here we likewise identify an N-glycosylated species in the RNA fraction. However, our results suggest that the reported RNase sensitivity of the glycosylated species depends on the specific RNA purification method. This suggests the possibility of copurifying unexpected RNase-insensitive N-glycoconjugates during glycoRNA isolation. The co-existence of two independent, yet highly similar molecular entities, complicates biochemical assays on glycoRNA and calls for more specific approaches for glycoRNA analysis. To address this, we propose a control experiment that can help distinguish genuine glycoRNA species from copurified glycoconjugates.
Authors
- Vered Raz (ORCID: https://orcid.org/0000-0003-3152-1952)
- Christian Büll (ORCID: https://orcid.org/0000-0001-7785-2920)
- Adam Pomorski (ORCID: https://orcid.org/0000-0002-0669-3421)
- Marvin E. Tanenbaum (ORCID: https://orcid.org/0000-0001-8762-0090)
- Kirsten Janssen
- Pascal Miesen (ORCID: https://orcid.org/0000-0002-6378-0871)
- Zeshi Li (ORCID: https://orcid.org/0000-0002-8358-3162)
- Bhagyashree S. Joshi (ORCID: https://orcid.org/0000-0002-9551-8640)
- Chirlmin Joo (ORCID: https://orcid.org/0000-0003-2803-0335)
- Sung‐Chul Kim (ORCID: https://orcid.org/0000-0003-0512-1154)
- Yong-geun Choi
- Jan-Willem H. Langenbach
- D. Brink (ORCID: https://orcid.org/0009-0004-0874-5573)
Institutions
- Leiden University (NL)
- Pohang University of Science and Technology (KR)
- Ewha Womans University (KR)
- Radboud University Nijmegen (NL)
- Utrecht University (NL)
- Yonsei University (KR)
- University of Wrocław (PL)
- Radboud University Medical Center (NL)
- University Medical Center Utrecht (NL)
- Institute for Basic Science (KR)
- Radboud Institute for Molecular Life Sciences (NL)
- Oncode Institute (NL)
- Pharmo Institute (NL)
- Whitehead Institute for Biomedical Research (US)
- Delft University of Technology (NL)
Publication Details
- Journal
- Experimental & Molecular Medicine
- Published
- 2025-11-15
- DOI
- https://doi.org/10.1038/s12276-025-01585-z
- Citations
- 2
- Primary Topic
- RNA and protein synthesis mechanisms
- Type
- article
- Field-Weighted Citation Impact
- 1.61
Funders
- National Research Foundation
- Universiteit Utrecht
- Technische Universiteit Delft
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek
- Institute for Basic Science
- Ministry of Science and ICT, South Korea