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Optimised detection of mitochondrial DNA strand breaks

Lookup NU author(s): Rebecca Hanna, David Moore, Professor Mark Birch-Machin

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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND).


Abstract

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 30-fold mtDNA damage in human skin cells induced by hydrogen peroxide and solar simulated radiation.


Publication metadata

Author(s): Hanna R, Crowther J, Bulsara P, Wang X, Moore D, Birch-Machin MA

Publication type: Article

Publication status: Published

Journal: Mitochondrion

Year: 2019

Volume: 46

Pages: 172-178

Online publication date: 04/05/2018

Acceptance date: 30/04/2018

Date deposited: 08/05/2018

ISSN (print): 1567-7249

ISSN (electronic): 1872-8278

Publisher: Elsevier

URL: https://doi.org/10.1016/j.mito.2018.04.009

DOI: 10.1016/j.mito.2018.04.009

Notes:


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