Mitochondrially targeted ZFNs for selective degradation of pathogenic mitochondrial genomes bearing large-scale deletions or point mutations.

TitleMitochondrially targeted ZFNs for selective degradation of pathogenic mitochondrial genomes bearing large-scale deletions or point mutations.
Publication TypeJournal Article
Year of Publication2014
AuthorsGammage, PA, Rorbach, J, Vincent, AI, Rebar, EJ, Minczuk, MA
JournalEMBO Mol Med
Volume6
Issue4
Pagination458-66
Date Published2014 Apr
ISSN1757-4684
KeywordsDeoxyribonucleases, DNA, Mitochondrial, Genetic Therapy, Genome, Mitochondrial, Humans, Mitochondria, Mitochondrial Diseases, Point Mutation, Protein Transport, Sequence Deletion
Abstract

We designed and engineered mitochondrially targeted obligate heterodimeric zinc finger nucleases (mtZFNs) for site-specific elimination of pathogenic human mitochondrial DNA (mtDNA). We used mtZFNs to target and cleave mtDNA harbouring the m.8993T>G point mutation associated with neuropathy, ataxia, retinitis pigmentosa (NARP) and the "common deletion" (CD), a 4977-bp repeat-flanked deletion associated with adult-onset chronic progressive external ophthalmoplegia and, less frequently, Kearns-Sayre and Pearson's marrow pancreas syndromes. Expression of mtZFNs led to a reduction in mutant mtDNA haplotype load, and subsequent repopulation of wild-type mtDNA restored mitochondrial respiratory function in a CD cybrid cell model. This study constitutes proof-of-principle that, through heteroplasmy manipulation, delivery of site-specific nuclease activity to mitochondria can alleviate a severe biochemical phenotype in primary mitochondrial disease arising from deleted mtDNA species.

DOI10.1002/emmm.201303672
Alternate JournalEMBO Mol Med
Citation Key10.1002/emmm.201303672
PubMed ID24567072
PubMed Central IDPMC3992073
Grant ListMC_U105697135 / / Medical Research Council / United Kingdom
/ / Medical Research Council / United Kingdom