title: Using engineered endonucleases to create knockout and knockin zebrafish models.
abstract: Over the last few years, the technology to create targeted knockout and knockin zebrafish animals has exploded. We have gained the ability to create targeted knockouts through the use of zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats/CRISPR associated system (CRISPR/Cas). Furthermore, using the high-efficiency TALEN system, we were able to create knockin zebrafish using a single-stranded DNA (ssDNA) protocol described here. Through the use of these technologies, the zebrafish has become a valuable vertebrate model and an excellent bridge between the invertebrate and mammalian model systems for the study of human disease.
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic, Guggenheim 1321A, 200 1st Street South, Rochester, MN, 55905, USA.
authors.forename: Victoria M
authors.initials: VM
authors.lastname: Bedell
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Ekker
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004260:DNA Repair; D004277:DNA, Single-Stranded; D004720:Endonucleases / Q000235:genetics* / Q000378:metabolism*; D055879:Gene Knock-In Techniques; D055786:Gene Knockout Techniques; D005818:Genetic Engineering / Q000379:methods*; D005820:Genetic Testing / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D005854:Germ Cells / Q000378:metabolism; D059765:Homologous Recombination; D023421:Models, Animal; D015027:Zebrafish / Q000235:genetics*
chemical_list: D004277:DNA, Single-Stranded; D004720:Endonucleases