Molecular Biotechnology

, Volume 19, Issue 3, pp 297–304

Contemporary gene targeting strategies for the novice

Authors

  • Siew-Sim Cheah
    • Department of Molecular GeneticsThe University of Texas M. D. Anderson Cancer Center
    • Department of Molecular GeneticsThe University of Texas M. D. Anderson Cancer Center
Protocol

DOI: 10.1385/MB:19:3:297

Cite this article as:
Cheah, S. & Behringer, R.R. Mol Biotechnol (2001) 19: 297. doi:10.1385/MB:19:3:297

Abstract

Gene targeting in mouse embryonic stem (ES) cells is a fundamental methodology for generating mice with precise genetic modifications. Although there are many complex gene targeting strategies for creating a variety of diverse mutations in mice, most investigators initially choose to generate a null allele. Here we provide a guide for the novice to generate a null allele for a protein coding gene using a fundamental gene targeting strategy. Ultimately, a well considered gene targeting strategy saves significant amounts of time, money, and research animal lives. The straightforward strategy presented here bypasses many of the pitfalls associated with gene knockouts generated by novices. This guide also serves as a foundation for subsequently designing more complex gene targeting strategies.

Index Entries

Gene targetingembryonic stem cellsCre/loxPknockouthomologous recombination
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Copyright information

© Humana Press Inc 2001