UNIT 4.59 Synthesis of Acetylene-Substituted Probes with Benzene-Phosphate Backbones for RNA Labeling
Published Online: 24 JUN 2014
Copyright © 2013 John Wiley & Sons, Inc. All rights reserved.
Lab Protocol Title
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.
- Published Online: 24 JUN 2014
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.
- RNA labeling;
- Huisgen cycloaddition;
- click chemistry