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

Imidazole, ACS reagent, ≥99% (titration)

Sigma-Aldrich

Imidazole, ≥99% (titration), crystalline

Sigma-Aldrich

Water, PCR Reagent

Sigma-Aldrich

Water, Nuclease-Free Water, for Molecular Biology

Sigma-Aldrich

E-Toxate^™ Water, endotoxin, free

Sigma-Aldrich

Water, for embryo transfer, sterile-filtered, BioXtra, suitable for mouse embryo cell culture

Sigma-Aldrich

Trichloroacetic acid, suitable for electrophoresis, suitable for fixing solution (for IEF and PAGE gels), ≥99%

Sigma-Aldrich

Trichloroacetic acid, BioXtra, ≥99.0%

Sigma-Aldrich

Triethylamine, ≥99%

Sigma-Aldrich

Imidazole, for molecular biology, ≥99% (titration)

Sigma-Aldrich

Triethylamine, ≥99.5%

Sigma-Aldrich

Triethylamine, for protein sequence analysis, ampule, ≥99.5% (GC)

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Water, for cell biology, sterile ultrafiltered

Sigma-Aldrich

Water, for molecular biology, sterile filtered

Supelco

Water, for HPCE, for luminescence, suitable for UV/Vis spectroscopy

Sigma-Aldrich

Triethylamine, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, for the determination of Fe in blood according to Heilmeyer, ≥99.5%

Sigma-Aldrich

Imidazole, BioUltra, for molecular biology, ≥99.5% (GC)

Sigma-Aldrich

Imidazole, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Imidazole, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

Trichloroacetic acid, BioUltra, ≥99.5% (T)

Sigma-Aldrich

Azidomethyl phenyl sulfide, 95%

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

Sigma-Aldrich

2,6-Lutidine

Sigma-Aldrich

2,6-Lutidine, ReagentPlus^®, 98%

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol