Simple and Stereoselective Preparation of an 4‐(Aminomethyl)‐1,2,3‐triazolyl Nucleoside Phosphoramidite

Abstract

A simple and stereoselective synthesis of a protected 4-(aminomethyl)-1-(2-deoxy-β-D-ribofuranosyl)-1,2,3-triazole cyanoethyl phosphoramidite was developed for the modification of synthetic oligonucleotides. The configuration of the 1,2,3-triazolyl moiety with respect to the deoxyribose was unambiguously determined in ROESY experiments. The aminomethyl group of the triazolyl nucleotide was fully functional in labelling reactions. Furthermore, the hybridization behavior of 5' triazole-terminated oligonucleotide was similar to that of 5' aminohexyl-terminated oligomer with the same sequence. Internal modifications of the oligonucleotide strands resulted in significant decrease of duplex stability.

Synthesis and Photophysical Properties of Triazolyl IrIII Nucleosides

“…The crude product was then separated by column chromatography on silica gel (dichloromethane/methanol 95:5, gradient until 92:8) and characterized. Adapted from literature …”

Mass-spectrometry analysis of modifications at DNA termini induced by DNA polymerases

“…The amino-modifier for labeling the 5′-end of the primer was synthesized from 5′ O -dimethoxytrityl thymidine by selective alkylation at N 3 as described in the Supplementary Information. Primer labeling was performed using activated Cy3 dye 48 following a published protocol 49 . The compositions of all synthesized oligonucleotides were confirmed by MALDI mass spectrometry analysis.…”

Targeting duplex DNA with chimeric α,β-triplex-forming oligonucleotides

“…Removal of the 5′- O -DMTr group was achieved by treatment with 2% aqueous TFA for 1 min followed by Et 3 N neutralization and precipitation with 2% LiClO 4 in acetone. Fluorescent labelling of oligonucleotides with the Cy3 dye was carried out as previously described (33). Oligonucleotide composition was confirmed by MALDI mass spectrometry using a Bruker Reflex IV mass spectrometer with hydroxypicolinic acid or 2-amino-5-nitropyridine as a matrix.…”