title: Generation of targeted mouse mutants by embryo microinjection of TALENs.
abstract: Gene engineering for generating targeted mouse mutants is a key technology for biomedical research. Using TALENs as nucleases to induce targeted double-strand breaks, the mouse genome can be directly modified in zygotes in a single step, without the need for embryonic stem cells. Thereby, knockout and knockin alleles can be generated fast and efficiently by embryo microinjection of TALEN mRNAs and targeting vectors. In this article we present an introduction into the TALEN technology and provide protocols for the application of TALENs in mouse zygotes.
authors.affiliation: Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Munich, Germany. Electronic address: benedikt.wefers@helmholtz-muenchen.de.
authors.forename: Benedikt
authors.initials: B
authors.lastname: Wefers
authors.affiliation: Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Munich, Germany. Electronic address: oskar.ortiz@helmholtz-muenchen.de.
authors.forename: Oskar
authors.initials: O
authors.lastname: Ortiz
authors.affiliation: Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Munich, Germany; Technische Universität München, 85350 Freising-Weihenstephan, Germany; Deutsches Zentrum für Neurodegenerative Erkrankungen e.V. (DZNE), 80336 Munich, Germany; Max Planck Institute of Psychiatry, 80804 Munich, Germany. Electronic address: wurst@helmholtz-muenchen.de.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wurst
authors.affiliation: Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Munich, Germany; Technische Universität München, 85350 Freising-Weihenstephan, Germany. Electronic address: ralf.kuehn@helmholtz-muenchen.de.
authors.forename: Ralf
authors.initials: R
authors.lastname: Kühn
mesh_terms: D000818:Animals; D003851:Deoxyribonucleases / Q000737:chemistry / Q000235:genetics; D055879:Gene Knock-In Techniques; D055786:Gene Knockout Techniques; D051379:Mice; D008845:Microinjections; D016297:Mutagenesis, Site-Directed / Q000379:methods*
chemical_list: D003851:Deoxyribonucleases
keywords: Gene targeting; Knockin; Knockout; Mouse; One-cell embryo; TALEN