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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

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

Supelco

Trichloroacetic acid, PESTANAL^®, analytical standard

Supelco

Water, for TOC analysis

Sigma-Aldrich

Methanol, NMR reference standard

Supelco

Density Standard 998 kg/m³, H&D Fitzgerald Ltd. Quality

Sigma-Aldrich

Water, BioPerformance Certified

Supelco

Water, ACS reagent, for ultratrace analysis

Sigma-Aldrich

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

Sigma-Aldrich

Trichloroacetic acid, biotech. grade, redistilled, ≥99%

Sigma-Aldrich

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

Supelco

Methanol, analytical standard

Sigma-Aldrich

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

Sigma-Aldrich

Imidazole, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

Imidazole, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

Sigma-Aldrich

Water, tested according to Ph. Eur.

Supelco

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

Supelco

2,6-Lutidine, analytical standard

Sigma-Aldrich

2,6-Lutidine

Sigma-Aldrich

Water, for cell biology, sterile ultrafiltered

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

2,6-Lutidine, ReagentPlus^®, 98%

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol

Sigma-Aldrich

Azidomethyl phenyl sulfide, 95%

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC

Sigma-Aldrich

Methanol, anhydrous, 99.8%