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Manipulating the Metazoan Mitochondrial Genome with Targeted Restriction Enzymes

Author:
Xu, Hong, DeLuca, Steven Z., O'Farrell, Patrick H.
Source:
Science 2008 v.321 no.5888 pp. 575-577
ISSN:
0036-8075
Subject:
Drosophila, cytochrome-c oxidase, genes, genetic techniques and protocols, genomics, germ cells, growth retardation, humans, longevity, male fertility, males, mitochondria, mitochondrial DNA, mitochondrial genome, models, muscular atrophy, mutants, mutation, neurodegenerative diseases, progeny, restriction endonucleases
Abstract:
High copy number and random segregation confound genetic analysis of the mitochondrial genome. We developed an efficient selection for heritable mitochondrial genome (mtDNA) mutations in Drosophila, thereby enhancing a metazoan model for study of mitochondrial genetics and mutations causing human mitochondrial disease. Targeting a restriction enzyme to mitochondria in the germline compromised fertility, but escaper progeny carried homoplasmic mtDNA mutations lacking the cleavage site. Among mutations eliminating a site in the cytochrome c oxidase gene, mt:CoIA³⁰²T was healthy, mt:CoIR³⁰¹L was male sterile but otherwise healthy, and mt:CoIR³⁰¹S exhibited a wide range of defects, including growth retardation, neurodegeneration, muscular atrophy, male sterility, and reduced life span. Thus, germline expression of mitochondrial restriction enzymes creates a powerful selection and has allowed direct isolation of mitochondrial mutants in a metazoan.
Agid:
2058035