A qPCR-based assay to quantify oxidized guanine and other FPG-sensitive base lesions within telomeric DNA

Nathan O'Callaghan, Natalie Baack, Razinah Sharif, Michael Fenech

Research output: Contribution to journalArticlepeer-review

29 Citations (Scopus)

Abstract

Telomere shortening is an important risk factor for cancer and accelerated aging. However, it is becoming evident that oxidatively damaged DNA within the telomere sequence may also cause telomere dysfunction. Here we describe a reliable, cost-effective quantitative PCR (qPCR)-based method to measure the amount of oxidized residues within telomeric DNA that are recognized and excised by formamidopyridine DNA glycosylase (FPG). We also report that in an in vitro model of oxidative stress oxidized base lesions measured using this method are more prevalent within telomeric sequences. Furthermore, this method is sufficiently sensitive to detect changes in oxidative stress induced by zinc deficiency and hydrogen peroxide within the physiological range.

Original languageEnglish
Pages (from-to)403-412
Number of pages10
JournalBioTechniques
Volume51
Issue number6
DOIs
Publication statusPublished or Issued - 1 Dec 2011

Keywords

  • Oxidative DNA damage
  • Oxoguanine
  • Telomere
  • qPCR

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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