Optimised detection of mitochondrial DNA strand breaks

被引：6

作者：

Hanna, Rebecca ^{[1
]}

Crowther, Jonathan M. ^{[2
]}

Bulsara, Pallav A. ^{[3
]}

Wang, Xuying ^{[4
]}

Moore, David J. ^{[3
]}

Birch-Machin, Mark A. ^{[1
]}

机构：

[1] Newcastle Univ, Med Sch, Inst Cellular Med, Dermatol Sci, Newcastle NE2 4HH, England

[2] JMC Sci Consulting Ltd, Egham TW208LL, Surrey, England

[3] GSK Consumer Healthcare, 184 Liberty Corner Rd, Warren, NJ 07059 USA

[4] GSK Consumer Healthcare, 1250 S Collegeville Rd, Collegeville, PA 19426 USA

来源：

MITOCHONDRION | 2019年 / 46卷

基金：

英国生物技术与生命科学研究理事会;

关键词：

Mitochondrial DNA damage; Biomarker; Oxidative stress; Long range qPCR; Human; Skin; DAMAGE; PCR;

D O I：

10.1016/j.mito.2018.04.009

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Intrinsic and extrinsic factors that induce cellular oxidative stress damage tissue integrity and promote ageing, resulting in accumulative strand breaks to the mitochondrial DNA (mtDNA) genome. Limited repair mechanisms and close proximity to superoxide generation make mtDNA a prominent biomarker of oxidative damage. Using human DNA we describe an optimised long-range qPCR methodology that sensitively detects mtDNA strand breaks relative to a suite of short mitochondrial and nuclear DNA housekeeping amplicons, which control for any variation in mtDNA copy number. An application is demonstrated by detecting 16-36-fold mtDNA damage in human skin cells induced by hydrogen peroxide and solar simulated radiation.

引用

页码：172 / 178

页数：7

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