title: Automated Chemical Solid-Phase Synthesis and Deprotection of 5-Hydroxymethylcytosine-Containing RNA.
abstract: 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.
authors.affiliation: Institute of Organic Chemistry and Center for Molecular Biosciences, CMBI, Leopold-Franzens University, Innrain 80-82, 6020, Innbruck, Austria.
authors.forename: Christian
authors.initials: C
authors.lastname: Riml
authors.affiliation: Institute of Organic Chemistry and Center for Molecular Biosciences, CMBI, Leopold-Franzens University, Innrain 80-82, 6020, Innbruck, Austria. ronald.micura@uibk.ac.at.
authors.forename: Ronald
authors.initials: R
authors.lastname: Micura
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives* / Q000737:chemistry; D044127:Epigenesis, Genetic; D008745:Methylation; D012313:RNA / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification; D060327:Solid-Phase Synthesis Techniques
chemical_list: C011865:5-hydroxymethylcytosine; D012313:RNA; D044503:5-Methylcytosine
keywords: Cytosine; Epigenetics; Methylation; RNA; Solid-phase synthesis