Unit

UNIT 23.5 Production of a Heterozygous Mutant Cell Line by Homologous Recombination (Single Knockout)

  1. Richard Mortensen

Published Online: 1 APR 2008

DOI: 10.1002/0471142727.mb2305s82

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Mortensen, R. 2008. Production of a Heterozygous Mutant Cell Line by Homologous Recombination (Single Knockout). Current Protocols in Molecular Biology. 82:23.5:23.5.1–23.5.11.

Author Information

  1. University of Michigan Medical School, Ann Arbor, Michigan

Publication History

  1. Published Online: 1 APR 2008
  2. Published Print: APR 2008

Abstract

Gene targeting by homologous recombination is a powerful and widely used technique for introduction of specific gene mutations (frequently a gene inactivation) in transgenic animals. The basic method detailed in this unit uses sequences homologous to the endogenous gene flanking the mutation. While methods using bacterial artificial chromosomes (BACs) and recombineering may be used, in most cases simpler bacterial plasmid clones with several kb of homology are sufficient. This protocol details the strategic factors in designing the constructs for selection and screening for homologous recombination. Curr. Protoc. Mol. Biol. 82:23.5.1-23.5.11. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • homologous recombination;
  • heterozygous mouse;
  • mutation;
  • single knockout