Unit

UNIT 5.14 Synthesis of G-N2-(CH2)3-N2-G Trimethylene DNA Interstrand Cross-Links

  1. Francesca Gruppi,
  2. Tracy L. Johnson Salyard,
  3. Carmelo J. Rizzo

Published Online: 26 MAR 2014

DOI: 10.1002/0471142700.nc0514s56

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Gruppi, F., Johnson Salyard, T. L. and Rizzo, C. J. 2014. Synthesis of G-N2-(CH2)3-N2-G Trimethylene DNA Interstrand Cross-Links. Current Protocols in Nucleic Acid Chemistry. 5:5.14:5.14.1–5.14.15.

Author Information

  1. Departments of Chemistry and Biochemistry, Center for Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University, Nashville, Tennessee

Publication History

  1. Published Online: 26 MAR 2014

Abstract

The synthesis of G-N2-(CH2)3-N2-G trimethylene DNA interstrand cross-links (ICLs) in a 5′-CG-3′ and 5′-GC-3′ sequence from oligodeoxynucleotides containing N2-(3-aminopropyl)-2′-deoxyguanosine and 2-fluoro-O6-(trimethylsilylethyl)inosine is presented. Automated solid-phase DNA synthesis was used for unmodified bases and modified nucleotides were incorporated via their corresponding phosphoramidite reagent by a manual coupling protocol. The preparation of the phosphoramidite reagents for incorporation of N2-(3-aminopropyl)-2′-deoxyguanosine is reported. The high-purity trimethylene DNA interstrand cross-link product is obtained through a nucleophilic aromatic substitution reaction between the N2-(3-aminopropyl)-2′-deoxyguanosine- and 2-fluoro-O6-(trimethylsilylethyl)inosine-containing oligodeoxynucleotides. Curr. Protoc. Nucleic Acid Chem. 56:5.14.1-5.14.15. © 2014 by John Wiley & Sons, Inc.

Keywords:

  • interstrand cross-links;
  • DNA;
  • N2-(3-aminopropyl)-2′-deoxyguanosine