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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

Dichloromethane, Selectophore^™, ≥99.5%

Sigma-Aldrich

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

Sigma-Aldrich

Citric acid, 99%

Supelco

Pyridine, analytical standard

Supelco

N,N-Dimethylacetamide, analytical standard

Sigma-Aldrich

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

Sigma-Aldrich

Imidazole buffer Solution, BioUltra, 1 M in H₂O

Sigma-Aldrich

Citric acid, ≥99.5%, FCC, FG

Sigma-Aldrich

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

Sigma-Aldrich

N,N-Dimethylacetamide-d₉, 99 atom % D

Sigma-Aldrich

Acetic anhydride, 99.5%

Sigma-Aldrich

Acetonitrile, ≥99.5%, ACS reagent

Sigma-Aldrich

Acetic anhydride, ACS reagent, ≥98.0%

Sigma-Aldrich

Pyridine, ≥99%

Sigma-Aldrich

2,6-Dimethylpyridine, ≥99%

Sigma-Aldrich

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

Sigma-Aldrich

Iodine, 99.999% trace metals basis

Sigma-Aldrich

2-Hydroxybutyric acid sodium salt, 97%

Sigma-Aldrich

Iodine, ≥99.99% trace metals basis

Sigma-Aldrich

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

Sigma-Aldrich

Acetic anhydride, ReagentPlus^®, ≥99%

Supelco

Citric acid, certified reference material, TraceCERT^®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland

Sigma-Aldrich

Citric acid, Vetec^™, reagent grade, 99%

Sigma-Aldrich

Acetonitrile, ReagentPlus^®, 99%

Sigma-Aldrich

Pyridine, ReagentPlus^®, ≥99%

Sigma-Aldrich

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

Supelco

Methanol solution, contains 0.10 % (v/v) formic acid, UHPLC, suitable for mass spectrometry (MS), ≥99.5%

Sigma-Aldrich

Pyridine, ACS reagent, ≥99.0%

Sigma-Aldrich

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

Supelco

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