Open AccessMitochondrially targeted ZFN s for selective degradation of pathogenic mitochondrial genomes bearing large‐scale deletions or point mutations
Author(s) -
Gammage Payam A,
Rorbach Joanna,
Vincent Anna I,
Rebar Edward J,
Minczuk Michal
Publication year - 2014
Publication title -
embo molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.923
H-Index - 107
eISSN - 1757-4684
pISSN - 1757-4676
DOI - 10.1002/emmm.201303672
Subject(s) - zinc finger nuclease , biology , mitochondrial dna , heteroplasmy , point mutation , genetics , microbiology and biotechnology , human mitochondrial genetics , mutation , gene , genome , genome editing
We designed and engineered mitochondrially targeted obligate heterodimeric zinc finger nucleases (mt ZFN s) for site‐specific elimination of pathogenic human mitochondrial DNA (mt DNA ). We used mt ZFN s to target and cleave mt DNA 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 mt ZFN s led to a reduction in mutant mt DNA haplotype load, and subsequent repopulation of wild‐type mt DNA 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 mt DNA species.