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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

Sigma-Aldrich

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

Supelco

Methanol, analytical standard

Sigma-Aldrich

Trichloroacetic acid, BioUltra, ≥99.5% (T)

Sigma-Aldrich

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

Sigma-Aldrich

2,6-Lutidine, ReagentPlus^®, 98%

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

Triethylamine, ≥99.5%

Sigma-Aldrich

Imidazole, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

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

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Imidazole buffer Solution, BioUltra, 1 M in H₂O

Sigma-Aldrich

Imidazole, ≥99% (titration), crystalline

Sigma-Aldrich

Imidazole, ACS reagent, ≥99% (titration)

Sigma-Aldrich

Imidazole, for molecular biology, ≥99% (titration)

Sigma-Aldrich

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

Sigma-Aldrich

Triethylamine, ≥99.5%

Sigma-Aldrich

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

Sigma-Aldrich

Triethylamine, ≥99%

Sigma-Aldrich

Methanol solution, NMR reference standard, 4% in methanol-d₄ (99.8 atom % D), NMR tube size 3 mm × 8 in.

Supelco

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

Sigma-Aldrich

Water, for cell biology, sterile ultrafiltered

Sigma-Aldrich

Water, for molecular biology, sterile filtered

Supelco

2,6-Lutidine, analytical standard

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

Sigma-Aldrich

2,6-Lutidine

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, ≥99.0%