Automated Chemical Solid-Phase Synthesis and Deprotection of 5-Hydroxymethylcytosine-Containing RNA

  • Christian Riml
  • Ronald MicuraEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1562)


5-Hydroxymethylcytosine is an epigenetic base modification that is part of the demethylation pathway of 5-methylcytosine in DNA. 5-Methylcytosine is iteratively oxidized to 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxycytosine by enzymes of the TET protein family. Since the discovery of 5-hydroxymethylcytosine also in RNA its role in regulatory processes and metabolism remains elusive. To gain more insight into the function of RNA containing 5-hydroxymethylcytidine, innovative and interdisciplinary approaches are required. In this context, synthetic oligoribonucleotides containing 5-hyroxymethylcytidine are an inevitable tool. Therefore, in this chapter, we present the efficient synthesis of RNA oligonucleotides containing 5-hydroxymethylcytosine by solid-phase synthesis. The incorporation of the modified cytosine derivative into RNA is compatible with standard phosphoramidite-based synthesis procedures of oligoribonucleotides.

Key words

Solid-phase synthesis RNA Methylation Epigenetics Cytosine 



We thank the Austrian Science Fund FWF (I1040 and P27947) for funding.


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© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Institute of Organic Chemistry and Center for Molecular Biosciences, CMBILeopold-Franzens UniversityInnbruckAustria

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