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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.

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Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%

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Metanolo, suitable for HPLC, ≥99.9%

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Acqua, suitable for HPLC

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Diclorometano, suitable for HPLC, ≥99.8%, contains amylene as stabilizer

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Metanolo, ACS reagent, ≥99.8%

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Acetonitrile, HPLC Plus, ≥99.9%

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Acqua, Nuclease-Free Water, for Molecular Biology

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Diclorometano, contains 40-150 ppm amylene as stabilizer, ACS reagent, ≥99.5%

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Metanolo, suitable for HPLC, gradient grade, ≥99.9%

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Piridina, ACS reagent, ≥99.0%

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Metanolo, HPLC Plus, ≥99.9%

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Diclorometano, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%, contains 50-150 ppm amylene as stabilizer

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Iodio, ACS reagent, ≥99.8%, solid

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

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Acqua, HPLC Plus

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N,N-dimetilacetammide, suitable for HPLC, ≥99.9%

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Acetonitrile, ACS reagent, ≥99.5%

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Imidazolo, ReagentPlus^®, 99%

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Acqua, Deionized

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Acetonitrile, for HPLC, for UV, ≥99.9% (GC)

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N,N-dimetilacetammide, puriss. p.a., ≥99.5% (GC)

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Piridina, suitable for HPLC, ≥99.9%

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Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%

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Metanolo, suitable for HPLC, gradient grade, suitable as ACS-grade LC reagent, ≥99.9%

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N,N-dimetilacetammide, ReagentPlus^®, 99%

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Acido citrico, meets analytical specification of Ph. Eur., BP, USP, E330, anhydrous, 99.5-100.5% (based on anhydrous substance)

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Metanolo, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)

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Diclorometano, puriss. p.a., ACS reagent, reag. ISO, ≥99.9% (GC)

Sigma-Aldrich

Diclorometano, ACS reagent, ≥99.5%, contains 40-150 ppm amylene as stabilizer

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N,N-dimetilacetammide, ReagentPlus^®, ≥99%