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

doi:10.2144/000113788

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

Nutrition, Diabetes And Gut Health

Research output: Contribution to journal › Article › peer-review

38 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 language	English
Pages (from-to)	403-412
Number of pages	10
Journal	BioTechniques
Volume	51
Issue number	6
DOIs	https://doi.org/10.2144/000113788
Publication status	Published or Issued - Dec 2011

Keywords

Oxidative DNA damage
Oxoguanine
Telomere
qPCR

ASJC Scopus subject areas

Biotechnology
Biochemistry, Genetics and Molecular Biology(all)

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10.2144/000113788

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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, Telomere, qPCR",

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AU - O'Callaghan, Nathan

AU - Baack, Natalie

AU - 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 - Telomere

KW - qPCR

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

Abstract

Keywords

ASJC Scopus subject areas

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