Rapid Directional Shift of Mitochondrial DNA Heteroplasmy in Animal Tissues by a Mitochondrially Targeted Restriction Endonuclease

Frequently, mtDNA with pathogenic mutations coexist with wild-type genomes (mtDNA heteroplasmy). Mitochondrial dysfunction and disease ensue only when the proportion of mutated mtDNAs is high, thus a reduction in this proportion should provide an effective therapy for these disorders. We developed a...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2005-10, Vol.102 (40), p.14392-14397
Main Authors: Bayona-Bafaluy, Maria Pilar, Blits, Bas, Battersby, Brendan J., Shoubridge, Eric A., Moraes, Carlos T., Caskey, C. Thomas
Format: Article
Language:English
Subjects:
Adenoviridae
Animals
Biological Sciences
Blotting, Southern
Cell Line
Cell lines
COS cells
DNA
DNA Restriction Enzymes - metabolism
DNA Restriction Enzymes - therapeutic use
DNA, Mitochondrial - genetics
Enzymes
Female
Gene Targeting - methods
Genetic mutation
Genetic Therapy - methods
Genetic vectors
Genetic Vectors - genetics
Genomics
Haplotypes
Haplotypes - genetics
Hepatocytes
Inductive reasoning
Mice
Mice, Inbred NZB
Mitochondria
Mitochondrial Diseases - genetics
Mitochondrial Diseases - therapy
Mitochondrial DNA
Mutation
Mutation - genetics
Polymorphism, Restriction Fragment Length
Tissues
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Summary:Frequently, mtDNA with pathogenic mutations coexist with wild-type genomes (mtDNA heteroplasmy). Mitochondrial dysfunction and disease ensue only when the proportion of mutated mtDNAs is high, thus a reduction in this proportion should provide an effective therapy for these disorders. We developed a system to decrease specific mtDNA haplotypes by expressing a mitochondrially targeted restriction endonuclease, ApaLI, in cells of heteroplasmic mice. These mice have two mtDNA haplotypes, of which only one contains an ApaLI site. After transfection of cultured hepatocytes with mitochondrially targeted ApaLI, we found a rapid, directional, and complete shift in mtDNA heteroplasmy (2-6 h). We tested the efficacy of this approach in vivo, by using recombinant viral vectors expressing the mitochondrially targeted ApaLI. We observed a significant shift in mtDNA heteroplasmy in muscle and brain transduced with recombinant viruses. This strategy could prevent disease onset or reverse clinical symptoms in patients harboring certain heteroplasmic pathogenic mutations in mtDNA.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0502896102