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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

Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid

Imidazole, European Pharmacopoeia (EP) Reference Standard

Sigma-Aldrich

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

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

Sigma-Aldrich

Azidomethyl phenyl sulfide, 95%

Sigma-Aldrich

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

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, ≥99.0%

Sigma-Aldrich

Imidazole, ACS reagent, ≥99% (titration)

Sigma-Aldrich

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%

Ondansetron impurity E, European Pharmacopoeia (EP) Reference Standard

Supelco

Residual Solvent - Acetonitrile(solution in DMSO), Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

Triethylamine, ≥99.5%

Sigma-Aldrich

Methanol, JIS special grade, ≥99.8%

Supelco

Water, for ultratrace analysis, ACS reagent

Sigma-Aldrich

2,6-Lutidine, ReagentPlus^®, 98%

Supelco

2,6-Lutidine, analytical standard

Sigma-Aldrich

Water, for cell biology, sterile ultrafiltered

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol

Sigma-Aldrich

2,6-Lutidine

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Sigma-Aldrich

Methanol, SAJ first grade, ≥99.5%

Sigma-Aldrich

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

Supelco

Methanol, analytical standard

Sigma-Aldrich

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

Sigma-Aldrich

Methanol, suitable for HPLC

Sigma-Aldrich

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