Protocol

Rat Genomics

Volume 597 of the series Methods in Molecular Biology pp 211-225

Date:

Generation of Gene-Specific Mutated Rats Using Zinc-Finger Nucleases

  • Aron M. GeurtsAffiliated withHuman and Molecular Genetics Center, Department of Physiology, Medical College of Wisconsin Email author 
  • , Gregory J. CostAffiliated withSangamo BioSciences, Inc.
  • , Séverine RémyAffiliated withINSERM, UMR643CHU Nantes, Institut de Transplantation et de Recherche en Transplantation, ITERTUniversité de Nantes, Faculté de Médecine
  • , Xiaoxia CuiAffiliated withSigma-Aldrich Biotechnology
  • , Laurent TessonAffiliated withINSERM, UMR643CHU Nantes, Institut de Transplantation et de Recherche en Transplantation, ITERTUniversité de Nantes, Faculté de Médecine
  • , Claire UsalAffiliated withINSERM, UMR643CHU Nantes, Institut de Transplantation et de Recherche en Transplantation, ITERTUniversité de Nantes, Faculté de Médecine
  • , Séverine MénoretAffiliated withINSERM, UMR643CHU Nantes, Institut de Transplantation et de Recherche en Transplantation, ITERTUniversité de Nantes, Faculté de Médecine
  • , Howard J. JacobAffiliated withHuman and Molecular Genetics Center, Department of Physiology, Medical College of Wisconsin
  • , Ignacio AnegonAffiliated withINSERM, UMR643CHU Nantes, Institut de Transplantation et de Recherche en Transplantation, ITERTUniversité de Nantes, Faculté de Médecine
    • , Roland BuelowAffiliated withOpen Monoclonal Technology, Inc.

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Abstract

The genetic dissection of physiological and pathological traits in laboratory model organisms is accelerated by the ability to engineer loss-of-function mutations at investigator-specified loci. This chapter describes the use of zinc-finger nucleases (ZFNs) for the targeted disruption of endogenous rat genes directly in the embryo. ZFNs can specifically disrupt target genes in cultured rat cells and in embryos from inbred and outbred strains, leading to permanently genetically modified animals. This technology allows for the rapid, targeted modification of the rat genome.

Key words

Zinc-finger nuclease ZFN Knockout Rat model QTL Gene targeting Congenic NHEJ