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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

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

Sigma-Aldrich

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

Sigma-Aldrich

Acetonitrile, electronic grade, 99.999% trace metals basis

Sigma-Aldrich

N,N-Dimethylacetamide-d₉, 99 atom % D

Sigma-Aldrich

Acetic anhydride, ReagentPlus^®, ≥99%

Sigma-Aldrich

Acetic anhydride, 99.5%

Supelco

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

Sigma-Aldrich

N,N-Dimethylacetamide, anhydrous, 99.8%

Sigma-Aldrich

Pyridine, anhydrous, 99.8%

Sigma-Aldrich

Iodine, 99.999% trace metals basis

Sigma-Aldrich

2,6-Dimethylpyridine, ≥99%

Sigma-Aldrich

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

Sigma-Aldrich

Acetic anhydride, ACS reagent, ≥98.0%

Sigma-Aldrich

Pyridine, ≥99%

Sigma-Aldrich

Trichloroacetic acid solution, 6.1 N

Sigma-Aldrich

Methanol-¹²C, 99.95 atom % ¹²C

Sigma-Aldrich

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

Supelco

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

Sigma-Aldrich

Dichloromethane, puriss., meets analytical specification of Ph. Eur., NF, ≥99% (GC)

Sigma-Aldrich

Acetonitrile, ≥99.5% (GC)

Sigma-Aldrich

Pyridine, ≥99.5% (GC)

Sigma-Aldrich

Acetonitrile, ReagentPlus^®, 99%

Supelco

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

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

Acetic anhydride, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99% (GC)

Sigma-Aldrich

Pyridine, ReagentPlus^®, ≥99%

Sigma-Aldrich

Pyridine, biotech. grade, ≥99.9%