Skip to Content

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

Pyridine, ReagentPlus^®, ≥99%

SAFC

N,N-Dimethylacetamide, Ph. Eur.

Dichloromethane, suitable for HPLC, ≥99.7%

Sigma-Aldrich

Acetonitrile, suitable for HPLC-GC, ≥99.8% (GC)

USP

Residual Solvent Class 2 - Acetonitrile, United States Pharmacopeia (USP) Reference Standard

Sigma-Aldrich

Iodine, ReagentPlus^®, 99.7% trace metals basis, beads, 1-3 mm

Sigma-Aldrich

Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)

Sigma-Aldrich

Iodine, flakes, ReagentPlus^®, ≥99%

Sigma-Aldrich

N,N-Dimethylacetamide, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

Iodine, puriss., ≥99.5% (RT), particles (round)

Sigma-Aldrich

Acetonitrile, ≥99.5% (GC)

Supelco

Iodine, ReagentPlus^®, ≥99.8% (titration)

Sigma-Aldrich

Trichloroacetic acid solution, 6.1 N

Sigma-Aldrich

Methanol-¹²C, 99.95 atom % ¹²C

Supelco

Acetic anhydride, for GC derivatization, LiChropur^™, ≥99.0%

Sigma-Aldrich

2,6-Dimethylpyridine, ≥99%

Sigma-Aldrich

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

Sigma-Aldrich

Pyridine, anhydrous, 99.8%

Sigma-Aldrich

N,N-Dimethylacetamide-d₉, 99 atom % D

Sigma-Aldrich

Methanol solution, NMR reference standard, 4% in methanol-d₄ (99.8 atom % D), NMR tube size 3 mm × 8 in.

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

Sigma-Aldrich

Methanol solution, (Methanol:Dimethyl sulfoxide 1:1 (v/v))

Sigma-Aldrich

Methanol solution, contains 0.50 % (v/v) triethylamine

Supelco

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

Sigma-Aldrich

2-Hydroxybutyric acid sodium salt, 97%

Sigma-Aldrich

Imidazole buffer Solution, BioUltra, 1 M in H₂O

Sigma-Aldrich

Iodine, ACS reagent, ≥99.8%, solid

USP

Methyl alcohol, United States Pharmacopeia (USP) Reference Standard

USP

Citric acid, United States Pharmacopeia (USP) Reference Standard

Sigma-Aldrich

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