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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

Acetic anhydride, Arxada quality, ≥99.5% (GC)

Sigma-Aldrich

Trichloroacetic acid solution, 6.1 N

Sigma-Aldrich

2-Hydroxybutyric acid sodium salt, 97%

Sigma-Aldrich

Citric acid, ACS reagent, ≥99.5%

Sigma-Aldrich

Iodine, 99.999% trace metals basis

Sigma-Aldrich

N,N-Dimethylacetamide, ReagentPlus^®, ≥99%

Sigma-Aldrich

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

Supelco

Citric acid, Anhydrous, Pharmaceutical Secondary Standard; Certified Reference Material

Supelco

Dichloromethane solution, contains 10 % (v/v) methanol

Sigma-Aldrich

Dichloromethane, suitable for HPLC, ≥99.9%, contains 40-150 ppm amylene as stabilizer

Sigma-Aldrich

N,N-Dimethylacetamide-d₉, 99 atom % D

Sigma-Aldrich

Acetic anhydride, 99.5%

Sigma-Aldrich

Citric acid, ≥99.5%, FCC, FG

Sigma-Aldrich

N,N-Dimethylacetamide, suitable for peptide synthesis, ≥99.8% (GC)

Sigma-Aldrich

Pyridine, ≥99%

Sigma-Aldrich

Acetic anhydride, ReagentPlus^®, ≥99%

Sigma-Aldrich

2,6-Dimethylpyridine, ≥99%

Sigma-Aldrich

Iodine, anhydrous, beads, −10 mesh, 99.999% trace metals basis

Sigma-Aldrich

Pyridine, anhydrous, 99.8%

Sigma-Aldrich

Acetonitrile, ≥99.5%, ACS reagent

Supelco

Dichloromethane, analytical standard

Sigma-Aldrich

Dichloromethane, anhydrous, ≥99.8%, contains 40-150 ppm amylene as stabilizer

Sigma-Aldrich

N,N-Dimethylacetamide, anhydrous, 99.8%

Sigma-Aldrich

Methylamine, ≥99.0%

Supelco

N,N-Dimethylacetamide, analytical standard

Supelco

Pyridine, analytical standard

Supelco

Acetonitrile, suitable for HPLC, gradient grade, ≥99.9% (GC)

Supelco

Acetonitrile, analytical standard

Supelco

Dichloromethane, Selectophore^™, ≥99.5%

Sigma-Aldrich

Iodine, ≥99.99% trace metals basis