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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)

Sigma-Aldrich

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

Sigma-Aldrich

Imidazole, puriss. p.a., ≥99.5% (GC)

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Imidazole, BioUltra, for molecular biology, ≥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

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

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Water, for molecular biology, sterile filtered

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Triethylamine, puriss. p.a., ≥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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Methylamine solution, 33 wt. % in absolute ethanol ((denatured with 1% toluene))

Supelco

Water, for ultratrace analysis, ACS reagent

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Water, BioPerformance Certified

Supelco

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

Supelco

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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Imidazole, ACS reagent, ≥99% (titration)

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Trichloroacetic acid, BioXtra, ≥99.0%

Sigma-Aldrich

Trichloroacetic acid, ≥99.0% (titration)

Sigma-Aldrich

Trichloroacetic acid, ACS reagent, ≥99.0%

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Water, sterile-filtered, BioReagent, suitable for cell culture

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

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Water, Nuclease-Free Water, for Molecular Biology

Sigma-Aldrich

Triethylamine, ≥99.5%