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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

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

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

Sigma-Aldrich

Triethylamine, ≥99.5%

Supelco

Water, ACS reagent, for ultratrace analysis

Supelco

Water, for TOC analysis

Sigma-Aldrich

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

Sigma-Aldrich

Methanol, purification grade, 99.8%

Supelco

Density Standard 998 kg/m³, H&D Fitzgerald Ltd. Quality

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Water, BioPerformance Certified

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Imidazole, BioUltra, ≥99.5% (GC)

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

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Trichloroacetic acid, BioUltra, ≥99.5% (T)

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Methylamine solution, 2.0 M in methanol

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2,6-Lutidine, ReagentPlus^®, 98%

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Azidomethyl phenyl sulfide, 95%

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Methanol, anhydrous, 99.8%

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Water, for cell biology, sterile ultrafiltered

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Water, Molecular Biology, sterile filtered

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Water, tested according to Ph. Eur.

Supelco

2,6-Lutidine, analytical standard

Sigma-Aldrich

Methylamine solution, 40 wt. % in H₂O

Sigma-Aldrich

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

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

Sigma-Aldrich

Triethylamine, BioUltra, ≥99.5% (GC)

Sigma-Aldrich

Imidazole, puriss. p.a., ≥99.5% (GC)

Sigma-Aldrich

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

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, ≥99.0%

Sigma-Aldrich

Trichloroacetic acid, ≥99.0% (titration)

Sigma-Aldrich

Trichloroacetic acid, BioXtra, ≥99.0%

Sigma-Aldrich

Imidazole, ACS reagent, ≥99% (titration)