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

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

Research output: Contribution to journalArticle

23 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 - Dec 2011
Externally publishedYes

Fingerprint

DNA Glycosylases
Oxidative stress
Guanine
Assays
Telomere
Polymerase Chain Reaction
DNA
Oxidative Stress
Hydrogen Peroxide
Telomere Shortening
Zinc
Aging of materials
Costs
Costs and Cost Analysis
Neoplasms

Keywords

  • Oxidative DNA damage
  • Oxoguanine
  • qPCR
  • Telomere

ASJC Scopus subject areas

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

Cite this

A qPCR-based assay to quantify oxidized guanine and other FPG-sensitive base lesions within telomeric DNA. / O'Callaghan, Nathan; Baack, Natalie; Sharif @ Mohd Sharif, Razinah; Fenech, Michael.

In: BioTechniques, Vol. 51, No. 6, 12.2011, p. 403-412.

Research output: Contribution to journalArticle

O'Callaghan, Nathan ; Baack, Natalie ; Sharif @ Mohd Sharif, Razinah ; Fenech, Michael. / A qPCR-based assay to quantify oxidized guanine and other FPG-sensitive base lesions within telomeric DNA. In: BioTechniques. 2011 ; Vol. 51, No. 6. pp. 403-412.
@article{7597d5dc24b449469c5c6858eaeb9a75,
title = "A qPCR-based assay to quantify oxidized guanine and other FPG-sensitive base lesions within telomeric DNA",
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.",
keywords = "Oxidative DNA damage, Oxoguanine, qPCR, Telomere",
author = "Nathan O'Callaghan and Natalie Baack and {Sharif @ Mohd Sharif}, Razinah and Michael Fenech",
year = "2011",
month = "12",
doi = "10.2144/000113788",
language = "English",
volume = "51",
pages = "403--412",
journal = "BioTechniques",
issn = "0736-6205",
publisher = "Eaton Publishing Company",
number = "6",

}

TY - JOUR

T1 - A qPCR-based assay to quantify oxidized guanine and other FPG-sensitive base lesions within telomeric DNA

AU - O'Callaghan, Nathan

AU - Baack, Natalie

AU - Sharif @ Mohd Sharif, Razinah

AU - Fenech, Michael

PY - 2011/12

Y1 - 2011/12

N2 - 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.

AB - 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.

KW - Oxidative DNA damage

KW - Oxoguanine

KW - qPCR

KW - Telomere

UR - http://www.scopus.com/inward/record.url?scp=83255193508&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=83255193508&partnerID=8YFLogxK

U2 - 10.2144/000113788

DO - 10.2144/000113788

M3 - Article

VL - 51

SP - 403

EP - 412

JO - BioTechniques

JF - BioTechniques

SN - 0736-6205

IS - 6

ER -