Issue 39, 2022

Robust synthesis of 2′-azido modified RNA from 2′-amino precursors by diazotransfer reaction

Abstract

Azides are versatile bioorthogonal reporter moieties that are commonly used for site-specific labeling and functionalization of RNA to probe its biology. The preparation of azido modified nucleic acids by solid-phase synthesis is problematic due to the inherent reactivity of P(III) species with azides according to the Staudinger reaction. Various strategies have been developed to bypass this limitation and are often time-consuming, low-yielding and labor-intensive. In particular, the synthesis of RNA with internal 2′-azido modifications is restricted to a single approach that employs P(V) chemistry instead of the widely used P(III) phosphoramidite chemistry. To fill this methodological gap, we present a novel convenient path toward 2′-azido RNA from readily accessible 2′-amino RNA through treatment with the diazotizing reagent fluorosulfuryl azide (FSO2N3). A diazotransfer reaction was established for oligoribonucleotides of different lengths and secondary structures. The robustness of the approach was further demonstrated for RNAs containing multiple 2′-azido moieties and for RNAs containing other sensitive modifications such as thiouridine or methylated nucleobases with a positive charge. The synthetic ease of generating 2′-azido RNA will pave the way for biotechnological applications, in particular for siRNA technologies and for referencing the growing number of RNA metabolic labeling approaches that rely on 2′-azido nucleosides.

Graphical abstract: Robust synthesis of 2′-azido modified RNA from 2′-amino precursors by diazotransfer reaction

Supplementary files

Article information

Article type
Paper
Submitted
27 Aug 2022
Accepted
21 Sep 2022
First published
29 Sep 2022
This article is Open Access
Creative Commons BY license

Org. Biomol. Chem., 2022,20, 7845-7850

Robust synthesis of 2′-azido modified RNA from 2′-amino precursors by diazotransfer reaction

S. Moreno, J. M. Ramos Pittol, M. Hartl and R. Micura, Org. Biomol. Chem., 2022, 20, 7845 DOI: 10.1039/D2OB01560A

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