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

Methylamine solution, 33 wt. % in absolute ethanol ((denatured with 1% toluene))

Sigma-Aldrich

Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC

Sigma-Aldrich

Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid

Sigma-Aldrich

Acetonitrile solution, contains 10.0% acetone, 0.05% formic acid, 40.0% 2-propanol

Supelco

Water, for TOC analysis

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

Benzyl azide solution, ~0.5 M in dichloromethane, ≥95.0% (HPLC)

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol

Sigma-Aldrich

Acetonitrile solution, contains 0.05 % (w/v) ammonium formate, 5 % (v/v) water, 0.1 % (v/v) formic acid, suitable for HPLC

Sigma-Aldrich

Triethylamine, ≥99.5%

Sigma-Aldrich

Acetonitrile solution, contains 0.1 % (v/v) formic acid, suitable for HPLC

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

2,6-Lutidine, ReagentPlus^®, 98%

Sigma-Aldrich

Water, for cell biology, sterile ultrafiltered

Sigma-Aldrich

Triethylamine, ampule, ≥99.5% (GC)

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Water, Molecular Biology, sterile filtered

Sigma-Aldrich

Triethylamine, for amino acid analysis, ≥99.5% (GC)

Sigma-Aldrich

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

Sigma-Aldrich

Azidomethyl phenyl sulfide, 95%

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

Supelco

2,6-Lutidine, analytical standard

Sigma-Aldrich

2,6-Lutidine

Sigma-Aldrich

Triethylamine, ≥99.5%

Supelco

Methanol, analytical standard

Sigma-Aldrich

Imidazole, BioUltra, Molecular Biology, ≥99.5% (GC)

Sigma-Aldrich

Imidazole, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Imidazole, puriss. p.a., ≥99.5% (GC)

Supelco

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