Unit

UNIT 23.13 Recombineering-Based Procedure for Creating Cre/loxP Conditional Knockouts in the Mouse

  1. Jason Bouvier1,2,
  2. Jr-Gang Cheng1

Published Online: 1 JAN 2009

DOI: 10.1002/0471142727.mb2313s85

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Bouvier, J. and Cheng, J.-G. 2009. Recombineering-Based Procedure for Creating Cre/loxP Conditional Knockouts in the Mouse. Current Protocols in Molecular Biology. 85:23.13:23.13.1–23.13.27.

Author Information

  1. 1

    UNC-Chapel Hill, Chapel Hill, North Carolina

  2. 2

    Tetracore, Inc., Rockville, Maryland

Publication History

  1. Published Online: 1 JAN 2009
  2. Published Print: JAN 2009

Abstract

Gene targeting in the mouse is an essential tool for studying gene function and creating models of human disease. The method described in this unit takes advantage of bacterial artificial chromosomes, Cre/loxP and FLPe/FRT systems, and recently evolved recombineering approaches to simplify the preparation of targeting constructs for generation of conditional knockout (CKO) animals. This method has been used to generate >30 CKO constructs, most of them successfully used to target mouse ES cells and establish mutant mice. Design and preparation of the CKO construct, as well as step-wise troubleshooting guidelines, are described in detail. Curr. Protoc. Mol. Biol. 85:23.13.1-23.13.27. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • gene targeting;
  • conditional knockout;
  • recombineering;
  • mouse;
  • Cre/loxP