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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)

Sigma-Aldrich

Methanol, JIS 300, ≥99.8%, for residue analysis

Sigma-Aldrich

Triethylamine, SAJ first grade, ≥98.0%

Sigma-Aldrich

Methanol, SAJ first grade, ≥99.5%

Sigma-Aldrich

Methanol, JIS special grade, ≥99.8%

Sigma-Aldrich

Methanol, SAJ special grade

Sigma-Aldrich

Methanol, suitable for HPLC

Supelco

Water, for TOC analysis

Sigma-Aldrich

Acetonitrile solution, contains 0.05 % (w/v) ammonium formate, 5 % (v/v) water, 0.1 % (v/v) formic acid, suitable for HPLC

Sigma-Aldrich

Methanol, NMR reference standard

Supelco

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

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Water, BioPerformance Certified

Supelco

Water, ACS reagent, for ultratrace analysis

Sigma-Aldrich

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

Sigma-Aldrich

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

Imidazole, European Pharmacopoeia (EP) Reference Standard

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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

Methanol, suitable for HPLC, gradient grade, 99.93%

Sigma-Aldrich

Methylamine solution, 2.0 M in THF