Site-specific analysis of ribosomal 2′O-methylation by quantitative reverse transcription PCR under low deoxynucleotide triphosphate concentrations

Daniela Barros-Silva, Johan Tsui, Carmen Jerónimo, Guido Jenster, Elena S. Martens-Uzunova*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Ribose 2′O-methylation (Nm, ribomethylation) is the most abundant RNA modification present in rRNA. It has been shown that alterations in ribosomal 2′O-methylation at individual Nm sites likely reflect regulated cellular processes. Although several analytical approaches for Nm detection and profiling have been developed, a simple and affordable method for the screening and measurement of individual Nm sites in large numbers of tissue samples is required to examine their potential for clinical translation. Here, we describe a new quantitative reverse transcription PCR-based method that can sensitively assess ribomethylation levels at specific rRNA sites at single-nucleotide resolution in low input amounts of total RNA.

Original languageEnglish
Pages (from-to)225-235
Number of pages11
JournalBioTechniques
Volume74
Issue number5
DOIs
Publication statusPublished - 1 May 2023

Bibliographical note

Funding Information:
This work was a collaborative effort under the frame of the European COST Action CA16120: EPITRAN. D Barros-Silva was supported by FCT–Fundação para a Ciência e Tecnologia (SFRH/BD/136007/2018). ES Martens-Uzunova was supported by the Dutch Science Organization Open Competition Domain Science (NWO ENW Open-XS), grant number: OCENW.XS22.1.114 and a EUR fellowship 2018-CvB/RE/PJ/RA00279215. The funders had no role in the design of the study and collection, analysis and interpretation of data and in writing the manuscript. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in, or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed above.

Funding Information:
Financial & competing interests disclosure This work was a collaborative effort under the frame of the European COST Action CA16120: EPITRAN. D Barros-Silva was supported by FCT-Fundação para a Ciência e Tecnologia (SFRH/BD/136007/2018). ES Martens-Uzunova was supported by the Dutch Science Organization Open Competition Domain Science (NWO ENW Open-XS), grant number: OCENW.XS22.1.114 and a EUR fellowship 2018- CvB/RE/PJ/RA00279215. The funders had no role in the design of the study and collection, analysis and interpretation of data and in writing the manuscript. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in, or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed above.

Publisher Copyright:
© 2023 Elena Martens-Uzunova.

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