Engineered mtZFNs for manipulation of human mitochondrial DNA heteroplasmy

Gammage, P. A. , Van Haute, L. and Minczuk, M. (2016) Engineered mtZFNs for manipulation of human mitochondrial DNA heteroplasmy. In: Mitochondrial DNA. Series: Methods in molecular biology, 1351 (1351). Humana Press: New York, pp. 145-162. ISBN 9781493930395 (doi: 10.1007/978-1-4939-3040-1_11)

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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.

Item Type:Book Sections
Glasgow Author(s) Enlighten ID:Gammage, Dr Payam
Authors: Gammage, P. A., Van Haute, L., and Minczuk, M.
College/School:College of Medical Veterinary and Life Sciences > School of Cancer Sciences
Publisher:Humana Press

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