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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, HPLC Plus, ≥99.9%, poly-coated bottles

Supelco

Water, for ultratrace analysis, ACS reagent

Supelco

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

Supelco

Methanol, Pharmaceutical Secondary Standard; Certified Reference Material

Imidazole, European Pharmacopoeia (EP) Reference Standard

Sigma-Aldrich

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

Pure Water Density Standard, UKAS ISO/IEC17025 and ISO Guide 34 certified, density: 0.9982 g/mL at 20 °C, density: 0.9970 g/mL at 25 °C

Pure Water Density Standard, UKAS ISO/IEC17025 and ISO Guide 34 certified, density: 0.9982 g/mL at 20 °C, density: 0.9970 g/mL at 25 °C

Sigma-Aldrich

Imidazole, anhydrous, free-flowing, Redi-Dri^™, ACS reagent, ≥99%

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

Methanol, suitable for HPLC, gradient grade, 99.93%

Sigma-Aldrich

Water, tested according to Ph. Eur.

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

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

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2,6-Lutidine, ReagentPlus^®, 98%

Sigma-Aldrich

2,6-Lutidine

Supelco

2,6-Lutidine, analytical standard

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

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Trichloroacetic acid, BioXtra, ≥99.0%

Sigma-Aldrich

Trichloroacetic acid, ≥99.0% (titration)

Sigma-Aldrich

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

Sigma-Aldrich

Azidomethyl phenyl sulfide, 95%

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, ≥99.0%

Sigma-Aldrich

Imidazole, ACS reagent, ≥99% (titration)

Sigma-Aldrich

Imidazole, ≥99% (titration), crystalline

Sigma-Aldrich

Imidazole, BioUltra, ≥99.5% (GC)

Supelco

Methanol, analytical standard

Sigma-Aldrich

Imidazole, puriss. p.a., ≥99.5% (GC)