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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

Triethylamine, BioUltra, ≥99.5% (GC)

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

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Imidazole, puriss. p.a., ≥99.5% (GC)

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Imidazole, BioUltra, ≥99.5% (GC)

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Imidazole, ReagentPlus^®, 99%

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2,6-Lutidine, ReagentPlus^®, 98%

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Azidomethyl phenyl sulfide, 95%

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Methylamine solution, 40 wt. % in H₂O

Supelco

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

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

Supelco

Water, for ultratrace analysis, ACS reagent

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2,6-Lutidine, analytical standard

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

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

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Triethylamine, ≥99%

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Imidazole, ≥99% (titration), crystalline

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Trichloroacetic acid, BioXtra, ≥99.0%

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Trichloroacetic acid, ≥99.0% (titration)

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Trichloroacetic acid, ACS reagent, ≥99.0%

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Triethylamine, ≥99.5%

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Imidazole, Pharmaceutical Secondary Standard; Certified Reference Material