Unit

UNIT 4.58 Synthesis of 8-Oxoguanosine Phosphoramidite and Its Incorporation into Oligoribonucleotides

  1. Yosuke Taniguchi,
  2. Yohei Koga,
  3. Shigeki Sasaki

Published Online: 26 MAR 2014

DOI: 10.1002/0471142700.nc0458s56

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Taniguchi, Y., Koga, Y. and Sasaki, S. 2014. Synthesis of 8-Oxoguanosine Phosphoramidite and Its Incorporation into Oligoribonucleotides. Current Protocols in Nucleic Acid Chemistry. 4:4.58:4.58.1–4.58.10.

Author Information

  1. Graduate School of Pharmaceutical Sciences, Kyushu University, Maidashi, Higashi-ku, Fukuoka, Japan

Publication History

  1. Published Online: 26 MAR 2014

Abstract

The detailed synthetic protocol for preparation of the phosphoramidite of an oxidatively damaged ribonucleotide, 8-oxoguanosine (8-oxo-G), and its incorporation into RNA are described. The O6- and N7-bisdiphenylcarbamoyl-protected 8-oxoguanosine phosphoramidite was synthesized as a new phosphoramidite precursor unit for the synthesis of RNA. It was successfully incorporated into the RNA sequences, and the synthesized RNAs were completely deprotected with 28% aqueous ammonia solution at 55°C for 24 hr. After purification using HPLC, they were identified by MALDI-TOF mass measurement. The base-pairing properties showed that 8-oxo-G forms base pairs not only with rC or dC in anti-conformation, but also with rA in syn conformation within the RNA duplexes or RNA/DNA heteroduplexes. Curr. Protoc. Nucleic Acid Chem. 56:4.58.1-4.58.10. © 2014 by John Wiley & Sons, Inc.

Keywords:

  • 8-oxoguanosine;
  • oxidatively damaged ribonucleotide;
  • base pairing