Efficient Access to 3′-Terminal Azide-Modified
RNA for Inverse Click-Labeling Patterns
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Abstract
Labeled RNA becomes increasingly
important for molecular diagnostics
and biophysical studies on RNA with its diverse interaction partners,
which range from small metabolites to large macromolecular assemblies,
such as the ribosome. Here, we introduce a fast synthesis path to
3′-terminal 2′-<i>O</i>-(2-azidoethyl) modified
oligoribonucleotides for subsequent bioconjugation, as exemplified
by fluorescent labeling via Click chemistry for an siRNA targeting
the brain acid-soluble protein 1 gene (<i>BASP1</i>). Importantly,
the functional group pattern is inverse to commonly encountered alkyne-functionalized
“click”-able RNA and offers increased flexibility with
respect to multiple and stepwise labeling of the same RNA molecule.
Additionally, our route opens up a minimal step synthesis of 2′-<i>O</i>-(2-aminoethyl) modified pyrimidine nucleoside phosphoramidites
which are of widespread use to generate amino-modified RNA for <i>N</i>-hydroxysuccinimide (NHS) ester-based conjugations