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Conditional Gene Targeting: A Refined Method for Genetic Studies in Neurosciences

  • Jane Beil
  • Isabelle M. Mansuy
  • Thorsten Buch
Protocol
Part of the Neuromethods book series (NM, volume 81)

Abstract

Conditional mutagenesis in mice is a key approach in neurosciences that makes it possible to investigate the functions of defined genes within certain neural subpopulations. The approach is based on the combination of transgenic expression of a recombinase with targeted candidate genes containing recognition sequences of this recombinase generated by gene targeting. Here we describe three major recombination systems, the Cre/loxP, Flp/FRT, and Dre/rox, and present a procedure that allows the generation of target alleles for these systems. We provide detailed protocols for the culture and transfection of embryonic stem (ES) cells and for the identification and confirmation of homologous recombinants. This chapter also provides an overview of recent developments such as gene targeting in oocytes and discusses future perspectives.

Key words

Gene targeting Recombination Conditional Cre/loxP Flp/FRT Mutant mice Targeting vector ES cells 

Notes

Acknowledgements

We thank Caroline Siegenthaler for assembling Table 1.

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Jane Beil
    • 1
  • Isabelle M. Mansuy
    • 2
  • Thorsten Buch
    • 1
  1. 1.Institute for Medical Microbiology, Immunology, and HygieneTechnische Universität MünchenMunichGermany
  2. 2.Brain Research InstituteUniversity of Zürich and Swiss Federal Institute of Technology ZürichZurichSwitzerland

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