Unit

UNIT 13.6 Chemical Phosphorylation of Deoxyribonucleosides and Thermolytic DNA Oligonucleotides

  1. Cristina Ausín,
  2. Andrzej Grajkowski,
  3. Jacek Cieślak,
  4. Serge L. Beaucage

Published Online: 1 OCT 2006

DOI: 10.1002/0471142700.nc1306s26

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Ausín, C., Grajkowski, A., Cieślak, J. and Beaucage, S. L. 2006. Chemical Phosphorylation of Deoxyribonucleosides and Thermolytic DNA Oligonucleotides. Current Protocols in Nucleic Acid Chemistry. 26:13.6:13.6.1–13.6.21.

Author Information

  1. Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, Maryland

Publication History

  1. Published Online: 1 OCT 2006
  2. Published Print: SEP 2006

Abstract

The phosphorylating reagent bis[S-(4,4′-dimethoxytrityl)-2-mercaptoethyl]-N,N-diisopropylphosphoramidite is prepared in three steps from commercial methyl thioglycolate and diisopropylphosphoramidous dichloride. The phosphorylating reagent has been used successfully in the solid-phase synthesis of deoxyribonucleoside 5′-/3′-phosphate or -thiophosphate monoesters and oligonucleotide 5′-phosphate/-thiophosphate monoesters. Bis[S-(4,4′-dimethoxytrityl)-2-mercaptoethyl]-N,N-diisopropylphosphoramidite has also been employed in the construction of a thermolytic dinucleotide prodrug model to evaluate the ability of the reagent to produce thermosentive oligonucleotide prodrugs under mild temperature conditions (∼25°C) for potential therapeutic applications.

Keywords:

  • bis[S-(4,4′-dimethoxytrityl)-2-mercaptoethyl]-N,N-diisopropylphosphoramidite;
  • phosphorylation;
  • thermolytic oligonucleotide prodrugs;
  • solid-phase oligonucleotide synthesis