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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

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

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

Methanol, suitable for HPLC

Supelco

Water, for TOC analysis

Sigma-Aldrich

Methanol, JIS special grade, ≥99.8%

Sigma-Aldrich

Methanol, SAJ first grade, ≥99.5%

Sigma-Aldrich

Methanol, JIS 300, ≥99.8%, for residue analysis

Supelco

Water, suitable for ion chromatography

Sigma-Aldrich

Methanol, NMR reference standard

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

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

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%

Sigma-Aldrich

Triethylamine, ≥99.5%

Sigma-Aldrich

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

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Sigma-Aldrich

2,6-Lutidine, ReagentPlus^®, 98%

Sigma-Aldrich

Methylamine solution, 2.0 M in THF

Sigma-Aldrich

Methylamine solution, 2.0 M in methanol

Sigma-Aldrich

Trichloroacetic acid, BioUltra, ≥99.5% (T)

Sigma-Aldrich

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

Sigma-Aldrich

Imidazole, BioUltra, ≥99.5% (GC)

Supelco

Methanol, analytical standard

Sigma-Aldrich

Imidazole, 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

Water, for molecular biology, sterile filtered

Sigma-Aldrich

Water, tested according to Ph. Eur.