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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

Sigma-Aldrich

Methanol, NMR reference standard

Supelco

Water, for TOC analysis

Supelco

Water, suitable for ion chromatography

Sigma-Aldrich

Trichloroacetic acid, BioUltra, ≥99.5% (T)

Sigma-Aldrich

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

Sigma-Aldrich

Imidazole, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Imidazole, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

Triethylamine, for protein sequence analysis, ampule, ≥99.5% (GC)

Sigma-Aldrich

Water, for molecular biology, sterile filtered

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Water, tested according to Ph. Eur.

Sigma-Aldrich

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

Sigma-Aldrich

Water, for cell biology, sterile ultrafiltered

Supelco

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

Sigma-Aldrich

2,6-Lutidine

Sigma-Aldrich

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

Sigma-Aldrich

Triethylamine, purum, ≥99% (GC)

Supelco

Methanol, analytical standard

Sigma-Aldrich

Imidazole, BioUltra, for molecular biology, ≥99.5% (GC)

Supelco

2,6-Lutidine, analytical standard

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Sigma-Aldrich

2,6-Lutidine, ReagentPlus^®, 98%

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

Water, deuterium-depleted, ≤1 ppm (Deuterium oxide)

Sigma-Aldrich

Imidazole, ACS reagent, ≥99% (titration)

Sigma-Aldrich

Imidazole, ≥99% (titration), crystalline

Sigma-Aldrich

Trichloroacetic acid, suitable for electrophoresis, suitable for fixing solution (for IEF and PAGE gels), ≥99%

Sigma-Aldrich

Trichloroacetic acid, BioXtra, ≥99.0%