title: CRISPR-Cas9 enables conditional mutagenesis of challenging loci.
abstract: The International Knockout Mouse Consortium (IKMC) has produced a genome-wide collection of 15,000 isogenic targeting vectors for conditional mutagenesis in C57BL/6N mice. Although most of the vectors have been used successfully in murine embryonic stem (ES) cells, there remain a set of nearly two thousand genes that have failed to target even after several attempts. Recent attention has turned to the use of new genome editing technology for the generation of mutant alleles in mice. Here, we demonstrate how Cas9-assisted targeting can be combined with the IKMC targeting vector resource to generate conditional alleles in genes that have previously eluded targeting using conventional methods.
authors: Institute of Developmental Genetics, Helmholtz Zentrum München, Ingolstädter Landstraße 1, 85764 Neuherberg/Munich, Germany.
authors: Joel A
authors: JA
authors: Schick
authors: Institute of Developmental Genetics, Helmholtz Zentrum München, Ingolstädter Landstraße 1, 85764 Neuherberg/Munich, Germany.
authors: Claudia
authors: C
authors: Seisenberger
authors: Institute of Developmental Genetics, Helmholtz Zentrum München, Ingolstädter Landstraße 1, 85764 Neuherberg/Munich, Germany.
authors: Joachim
authors: J
authors: Beig
authors: Institute of Developmental Genetics, Helmholtz Zentrum München, Ingolstädter Landstraße 1, 85764 Neuherberg/Munich, Germany.
authors: Antje
authors: A
authors: Bürger
authors: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
authors: Vivek
authors: V
authors: Iyer
authors: Institute of Developmental Genetics, Helmholtz Zentrum München, Ingolstädter Landstraße 1, 85764 Neuherberg/Munich, Germany.
authors: Viola
authors: V
authors: Maier
authors: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
authors: Sajith
authors: S
authors: Perera
authors: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.|AstraZeneca Darwin Building (Unit 310), Cambridge Science Park, Milton Road, Cambridge, CB4 0WG, United Kingdom.
authors: Barry
authors: B
authors: Rosen
authors: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
authors: William C
authors: WC
authors: Skarnes
authors: Institute of Developmental Genetics, Helmholtz Zentrum München, Ingolstädter Landstraße 1, 85764 Neuherberg/Munich, Germany.|Technische Universität München-Weihenstephan, c/o Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764 Neuherberg/Munich, Germany.|German Center for Neurodegenerative Diseases (DZNE), Site Munich, Feodor-Lynen-Strasse 17, 81377 Munich, Germany.|Munich Cluster for Systems Neurology (SyNergy), Adolf-Butenandt-Institut Ludwig-Maximilians-Universität München, Feodor-Lynen-Strasse 17, 81377 Munich, Germany.
authors: Wolfgang
authors: W
authors: Wurst
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D018390:Gene Targeting; D056426:Genetic Loci; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D016296:Mutagenesis / Q000235:genetics*