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High-throughput chemical modification of oligonucleotides for systematic structure-activity relationship evaluation.

High-throughput chemical modification of oligonucleotides for systematic structure-activity relationship evaluation.

Bioconjugate chemistry (2014-11-15)

Daniel Zewge, Francis Gosselin, Denise M Kenski, Jenny Li, Vasant Jadhav, Yu Yuan, Sandhya S Nerurkar, David M Tellers, W Michael Flanagan, Ian W Davies

ABSTRACT

Chemical modification of siRNA is achieved in a high-throughput manner (96-well plate format) by copper catalyzed azide-alkyne cycloadditions. This transformation can be performed in one synthetic operation at up to four positions with complete specificity, good yield, and acceptable purity. As demonstrated here, this approach extends the current synthetic options for oligonucleotide modifications and simultaneously facilitates the systematic, rapid biological evaluation of modified siRNA.

MATERIALS

Product Number

Brand

Product Description

Supelco

Water, suitable for ion chromatography

Supelco

Water, for TOC analysis

Sigma-Aldrich

Methanol, suitable for NMR (reference standard)

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

Methylamine solution, 33 wt. % in absolute ethanol ((denatured with 1% toluene))

Sigma-Aldrich

Triethylamine, ≥99.5%

Sigma-Aldrich

Benzyl azide solution, ~0.5 M in dichloromethane, ≥95.0% (HPLC)

Supelco

Water, suitable for HPCE, suitable for luminescence, suitable for UV/Vis spectroscopy

Supelco

2,6-Lutidine, analytical standard

Sigma-Aldrich

2,6-Lutidine

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, for the determination of Fe in blood according to Heilmeyer, ≥99.5%

Sigma-Aldrich

Trichloroacetic acid, BioUltra, ≥99.5% (T)

Sigma-Aldrich

Triethylamine, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

Triethylamine, ampule, ≥99.5% (GC)

Sigma-Aldrich

Triethylamine, for amino acid analysis, ≥99.5% (GC)

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Imidazole, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

Imidazole, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Imidazole, BioUltra, Molecular Biology, ≥99.5% (GC)

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol

Sigma-Aldrich

2,6-Lutidine, ReagentPlus^®, 98%

Sigma-Aldrich

Azidomethyl phenyl sulfide, 95%

Sigma-Aldrich

Water, tested according to Ph. Eur.

Sigma-Aldrich

Water, for cell biology, sterile ultrafiltered

Sigma-Aldrich

Water, Molecular Biology, sterile filtered

Sigma-Aldrich

Water, Nuclease-Free Water, for Molecular Biology

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, ≥99.0%