title: Covalent Chemical 5'-Functionalization of RNA with Diazo Reagents.
abstract: Functionalization of RNA at the 5'-terminus is important for analytical and therapeutic purposes. Currently, these RNAs are synthesized de novo starting with a chemically functionalized 5'-nucleotide, which is incorporated into RNA using chemical synthesis or biochemical techniques. Methods for direct chemical modification of native RNA would provide an attractive alternative but are currently underexplored. Herein, we report that diazo compounds can be used to selectively alkylate the 5'-phosphate of ribo(oligo)nucleotides to give RNA labelled through a native phosphate ester bond. We applied this method to functionalize oligonucleotides with biotin and an orthosteric inhibitor of the eukaryotic initiation factor 4E (eIF4E), an enzyme involved in mRNA recognition. The modified RNA binds to eIF4E, demonstrating the utility of this labelling technique to modulate biological activity of RNA. This method complements existing techniques and may be used to chemically introduce a broad range of functional handles at the 5'-end of RNA.
authors.affiliation: Novartis Institutes for BioMedical Research, 181 Massachusetts Avenue, Cambridge, MA, 02139, USA. christian.gampe@novartis.com.
authors.forename: Christian M
authors.initials: CM
authors.lastname: Gampe
authors.affiliation: Novartis Institutes for BioMedical Research, 181 Massachusetts Avenue, Cambridge, MA, 02139, USA.
authors.forename: Micah
authors.initials: M
authors.lastname: Hollis-Symynkywicz
authors.affiliation: Novartis Institutes for BioMedical Research, 181 Massachusetts Avenue, Cambridge, MA, 02139, USA.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Zécri
mesh_terms: D001391:Azo Compounds / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D015394:Molecular Structure; D012313:RNA / Q000737:chemistry*
chemical_list: D001391:Azo Compounds; D012313:RNA
keywords: RNA; diazo compounds; labelling; nucleotides; phosphates