UNIT 4.30 Production of a Heterozygous Mutant Cell Line by Homologous Recombination (Single Knockout)

  1. Richard Mortensen

Published Online: 1 APR 2011

DOI: 10.1002/0471142301.ns0430s55

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Mortensen, R. 2011. Production of a Heterozygous Mutant Cell Line by Homologous Recombination (Single Knockout). Current Protocols in Neuroscience. 55:4.30:4.30.1–4.30.12.

Author Information

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

Publication History

  1. Published Online: 1 APR 2011
  2. Published Print: APR 2011


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. Neurosci. 55:4.30.1-4.30.12. © 2011 by John Wiley & Sons, Inc.


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