Unit

UNIT 4.59 Synthesis of Acetylene-Substituted Probes with Benzene-Phosphate Backbones for RNA Labeling

  1. Yoshiaki Kitamura1,
  2. Yoshihito Ueno2,
  3. Yukio Kitade1

Published Online: 24 JUN 2014

DOI: 10.1002/0471142700.nc0459s57

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Kitamura, Y., Ueno, Y. and Kitade, Y. 2014. Synthesis of Acetylene-Substituted Probes with Benzene-Phosphate Backbones for RNA Labeling. Current Protocols in Nucleic Acid Chemistry. 57:4.59:4.59.1–4.59.15.

Author Information

  1. 1

    Department of Chemistry and Biomolecular Science, Faculty of Engineering, Gifu University, Gifu, Japan

  2. 2

    Course of Applied Life Science, Faculty of Applied Biological Sciences, Gifu University, Gifu, Japan

Publication History

  1. Published Online: 24 JUN 2014

Abstract

Conversion of dimethyl 5-aminoisophthalate into the iodoarene via the corresponding diazonium intermediate, followed by Sonogashira coupling with trimethylsilylacetylene afford the alkynylarene, which is reduced with LiAlH4 to give 5-ethynyl-1,3-benzenedimethanol (BE). One hydroxyl group is protected with a 4,4′-dimethoxytrityl (DMTr) group and subsequently another hydroxyl group is phosphitylated to produce the phosphoramidite. The mono-DMTr compound is also modified to afford the corresponding succinate, which is then reacted with controlled pore glass (CPG) to provide the solid support. Either the phosphoramidite or the solid support is employed in solid-phase synthesis of RNA containing BE. RNA oligomers bearing BE rapidly react with 4-fluorobenzylazide to produce the cycloaddition products in good to excellent yield. Curr. Protoc. Nucleic Acid Chem. 57:4.59.1-4.59.15. © 2014 by John Wiley & Sons, Inc.

Keywords:

  • alkynes;
  • RNA labeling;
  • Huisgen cycloaddition;
  • click chemistry