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Engineered mtZFNs for Manipulation of Human Mitochondrial DNA Heteroplasmy

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Mitochondrial DNA

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1351))

Abstract

Enrichment of desired mitochondrial DNA (mtDNA) haplotypes, in both experimental systems and the clinic, is an end sought by many. Through use of a designer nuclease platform optimized for delivery to mitochondria—the mitochondrially targeted zinc finger-nuclease (mtZFN)—it is possible to discriminate between mtDNA haplotypes with specificity to the order of a single nucleotide substitution. Site-specific cleavage of DNA produces a shift in the heteroplasmic ratio in favor of the untargeted haplotype. Here, we describe protocols for assembly of paired, conventional tail–tail mtZFN constructs and experimental approaches to assess mtZFN activity in mammalian cell cultures.

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Authors and Affiliations

  1. MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Hills Road, Cambridge, CB2 0XY, UK

    Payam A. Gammage, Lindsey Van Haute & Michal Minczuk

Authors
  1. Payam A. Gammage
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  2. Lindsey Van Haute
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  3. Michal Minczuk
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Corresponding authors

Correspondence to Payam A. Gammage or Michal Minczuk .

Editor information

Editors and Affiliations

  1. Hudson Institute of Medical Research, Clayton, Melbourne, Victoria, Australia

    Matthew McKenzie

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© 2016 Springer Science+Business Media New York

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Gammage, P.A., Van Haute, L., Minczuk, M. (2016). Engineered mtZFNs for Manipulation of Human Mitochondrial DNA Heteroplasmy. In: McKenzie, M. (eds) Mitochondrial DNA. Methods in Molecular Biology, vol 1351. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3040-1_11

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  • DOI: https://doi.org/10.1007/978-1-4939-3040-1_11

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3039-5

  • Online ISBN: 978-1-4939-3040-1

  • eBook Packages: Springer Protocols

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