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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

Imidazole, ≥99% (titration), crystalline

Sigma-Aldrich

Imidazole, ACS reagent, ≥99% (titration)

Sigma-Aldrich

Trichloroacetic acid, ≥99.0% (titration)

Sigma-Aldrich

Trichloroacetic acid, BioXtra, ≥99.0%

Sigma-Aldrich

Water, for embryo transfer, sterile-filtered, BioXtra, suitable for mouse embryo cell culture

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, ≥99.0%

Sigma-Aldrich

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

Sigma-Aldrich

Triethylamine, ≥99%

Sigma-Aldrich

Triethylamine, ≥99.5%

Supelco

Water, suitable for ion chromatography

Sigma-Aldrich

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

Supelco

Water, for TOC analysis

Supelco

Water, for ultratrace analysis, ACS reagent

Sigma-Aldrich

Trichloroacetic acid, BioUltra, ≥99.5% (T)

Supelco

Methanol, analytical standard

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol

Sigma-Aldrich

Azidomethyl phenyl sulfide, 95%

Sigma-Aldrich

Imidazole, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Imidazole, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

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

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Water, tested according to Ph. Eur.

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

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

Sigma-Aldrich

Water, for cell biology, sterile ultrafiltered

Supelco

2,6-Lutidine, analytical standard

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

Sigma-Aldrich

2,6-Lutidine

Sigma-Aldrich

Methanol, anhydrous, 99.8%