UNIT 23.13 Recombineering-Based Procedure for Creating Cre/loxP Conditional Knockouts in the Mouse
Published Online: 1 JAN 2009
Copyright © 2009 by John Wiley & Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 JAN 2009
- Published Print: JAN 2009
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.
- gene targeting;
- conditional knockout;