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Mitochondrial transfection for studying organellar DNA repair, genome maintenance and aging

Lookup NU author(s): Dr Pierre Boesch, Professor Robert Lightowlers

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Abstract

Maintenance of the mitochondrial genome is a major challenge for cells, particularly as they begin to age. Although it is established that organelles possess regular DNA repair pathways, many aspects of these complex processes and of their regulation remain to be investigated. Mitochondrial transfection of isolated organelles and in whole cells with customized DNA synthesized to contain defined lesions has wide prospects for deciphering repair mechanisms in a physiological context. We document here the strategies currently developed to transfer DNA of interest into mitochondria. Methodologies with isolated mitochondria claim to exploit the protein import pathway or the natural competence of the organelles, to permeate the membranes or to use conjugal transfer from bacteria. Besides biolistics, which remains restricted to yeast and Chlamydomonas reinhardtii, nanocarriers or fusion proteins have been explored as methods to target custom DNA into mitochondria in intact cells. In further approaches, whole mitochondria have been transferred into recipient cells. Repair failure or error-prone repair leads to mutations which potentially could be rescued by allotopic expression of proteins. The relevance of the different approaches for the analysis of mitochondrial DNA repair mechanisms and of aging is discussed. (C) 2011 Elsevier Ireland Ltd. All rights reserved.


Publication metadata

Author(s): Mileshina D, Ibrahim N, Boesch P, Lightowlers RN, Dietrich A, Weber-Lotfi F

Publication type: Conference Proceedings (inc. Abstract)

Publication status: Published

Conference Name: Mechanisms of Ageing and Development: 3rd International Genome Dynamics in Neuroscience Conference on DNA Repair and Neurological Disease

Year of Conference: 2011

Pages: 412-423

ISSN: 0047-6374

Publisher: Elsevier Ireland Ltd

URL: http://dx.doi.org/10.1016/j.mad.2011.05.002

DOI: 10.1016/j.mad.2011.05.002

Library holdings: Search Newcastle University Library for this item

ISBN: 18726216


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