Manipulating mitochondrial DNA heteroplasmy by a mitochondrially targeted restriction endonuclease

Sarika Srivastava, Carlos T Moraes

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

Mutations in the mitochondrial DNA (mtDNA) can cause a variety of human diseases. In most cases, such mutations are heteroplasmic (i.e. mutated and wild-type mtDNA coexist) and a small percentage of wild-type sequences can have a strong protective effect against a metabolic defect. Because a genetic approach to correct mtDNA mutations is not currently available, the ability to modulate heteroplasmy would have a major impact in the phenotype of many patients with mitochondrial disorders. We show here that a restriction endonuclease targeted to mitochondria has this ability. A mitochondrially targeted Pstl degraded mtDNA harboring Pstl sites, in some cases leading to a complete loss of mitochondrial genomes. Recombination between DNA ends released by Pstl was not observed. When expressed in a heteroplasmic rodent cell line, containing one mtDNA haplotype with two sites for Pstl and another haplotype having none, the mitochondrial Pstl caused a significant shift in heteroplasmy, with an accumulation of the mtDNA haplotype lacking Pstl sites. These experiments provide proof of the principle that restriction endonucleases are feasible tools for genetic therapy of a sub-group of mitochondrial disorders. Although this approach is limited by the presence of mutation-specific restriction sites, patients with neuropathy, ataxia and retinitis pigmentosa (NARP) could benefit from it, as the T8399G mutation creates a unique restriction site that is not present in wild-type human mitochondrial DNA.

Original languageEnglish
Pages (from-to)3093-3099
Number of pages7
JournalHuman Molecular Genetics
Volume10
Issue number26
StatePublished - Dec 15 2001

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DNA Restriction Enzymes
Mitochondrial DNA
Mutation
Haplotypes
Mitochondrial Diseases
Mitochondrial Genome
Genetic Therapy
Genetic Recombination
Rodentia
Mitochondria
Phenotype
Cell Line
DNA

ASJC Scopus subject areas

  • Genetics

Cite this

Manipulating mitochondrial DNA heteroplasmy by a mitochondrially targeted restriction endonuclease. / Srivastava, Sarika; Moraes, Carlos T.

In: Human Molecular Genetics, Vol. 10, No. 26, 15.12.2001, p. 3093-3099.

Research output: Contribution to journalArticle

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