Abstract
We have established a reverse genetics approach for the routine generation of medaka (Oryzias latipes) gene knockouts. A cryopreserved library of N-ethyl-N-nitrosourea (ENU) mutagenized fish was screened by high-throughput resequencing for induced point mutations. Nonsense and splice site mutations were retrieved for the Blm, Sirt1, Parkin and p53 genes and functional characterization of p53 mutants indicated a complete knockout of p53 function. The current cryopreserved resource is expected to contain knockouts for most medaka genes.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphatases / genetics
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Animals
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Animals, Genetically Modified*
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Base Sequence
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DNA Helicases / genetics
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DNA Primers
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Ethylnitrosourea / administration & dosage
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Female
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Genes, p53
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Male
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Models, Genetic*
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Mutagenesis, Site-Directed
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Oryzias / genetics*
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Point Mutation
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RecQ Helicases
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Ubiquitin-Protein Ligases / genetics
Substances
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DNA Primers
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Ubiquitin-Protein Ligases
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parkin protein
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Adenosine Triphosphatases
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Bloom syndrome protein
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DNA Helicases
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RecQ Helicases
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Ethylnitrosourea