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

PMID25398098

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

02575

Supelco

Dichloromethane, analytical standard

91204

Supelco

Acetic anhydride, for GC derivatization, LiChropur^™, ≥99.0%

1424109

USP

Methyl alcohol, United States Pharmacopeia (USP) Reference Standard

1134368

USP

Citric acid, United States Pharmacopeia (USP) Reference Standard

650188

Sigma-Aldrich

Methanol solution, (Methanol:Dimethyl sulfoxide 1:1 (v/v))

691275

Sigma-Aldrich

Acetic anhydride, Arxada quality, ≥99.5% (GC)

791725

Sigma-Aldrich

Citric acid, anhydrous, free-flowing, Redi-Dri^™, ACS reagent, ≥99.5%

110086

Sigma-Aldrich

Acetonitrile, ReagentPlus^®, 99%

I3380

Supelco

Iodine, ReagentPlus^®, ≥99.8% (titration)

207772

Sigma-Aldrich

Iodine, ACS reagent, ≥99.8%, solid

154806

Sigma-Aldrich

N,N-Dimethylacetamide, spectrophotometric grade, ≥99%

P57506

Sigma-Aldrich

Pyridine, ReagentPlus^®, ≥99%

D65100

Sigma-Aldrich

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

D137510

Sigma-Aldrich

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

27109

Sigma-Aldrich

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

320269

Sigma-Aldrich

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

32222

Sigma-Aldrich

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

33553

Sigma-Aldrich

Pyridine, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.5% (GC)

676853

Sigma-Aldrich

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

33214

Sigma-Aldrich

Acetic anhydride, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99% (GC)

320498

Sigma-Aldrich

Pyridine, ReagentPlus^®, ≥99%

494410

Sigma-Aldrich

Pyridine, biotech. grade, ≥99.9%

376558

Sigma-Aldrich

Iodine, flakes, ReagentPlus^®, ≥99%

38840

Sigma-Aldrich

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

360570

Sigma-Aldrich

Pyridine, ACS reagent, ≥99.0%

34888

Sigma-Aldrich

Acetonitrile, ≥99.9% (GC)

34442

Sigma-Aldrich

Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)

60004

Sigma-Aldrich

Acetonitrile, ≥99.5% (GC)

03002

Sigma-Aldrich

Iodine, puriss., meets analytical specification of Ph. Eur., BP, USP, 99.8-100.5%

494445

Sigma-Aldrich

Acetonitrile, biotech. grade, ≥99.93%