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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

Supelco

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

USP

Citric acid, United States Pharmacopeia (USP) Reference Standard

Sigma-Aldrich

Acetonitrile, electronic grade, 99.999% trace metals basis

Sigma-Aldrich

Methanol-¹²C, 99.95 atom % ¹²C

Sigma-Aldrich

Trichloroacetic acid solution, 6.1 N

Sigma-Aldrich

Citric acid, ACS reagent, ≥99.5%

Supelco

Dichloromethane, Selectophore^™, for ion-selective electrodes, ≥99.5%

Supelco

Dichloromethane, analytical standard

Sigma-Aldrich

Citric acid, ≥99.5%, FCC, FG

Sigma-Aldrich

Pyridine, anhydrous, 99.8%

Sigma-Aldrich

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

Sigma-Aldrich

Imidazole buffer Solution, BioUltra, 1 M in H₂O

Sigma-Aldrich

Acetic anhydride, ReagentPlus^®, ≥99%

Sigma-Aldrich

2,6-Dimethylpyridine, ≥99%

Sigma-Aldrich

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

Supelco

N,N-Dimethylacetamide, analytical standard

Sigma-Aldrich

Citric acid, BioUltra, anhydrous, ≥99.5% (T)

Sigma-Aldrich

Acetic anhydride, ACS reagent, ≥98.0%

Sigma-Aldrich

Iodine, ≥99.99% trace metals basis

Sigma-Aldrich

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

Sigma-Aldrich

Citric acid, 99%

Citric acid, anhydrous, European Pharmacopoeia (EP) Reference Standard

USP

Methyl alcohol, United States Pharmacopeia (USP) Reference Standard

Supelco

Pyridine, Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

N,N-Dimethylacetamide-d₉, 99 atom % D

Sigma-Aldrich

2-Hydroxybutyric acid sodium salt, 97%

Sigma-Aldrich

Pyridine, ≥99%

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

Dichloromethane, ACS reagent, ≥99.5%, contains 40-150 ppm amylene as stabilizer