Production of a heterozygous mutant cell line by homologous recombination (single knockout)

Current Protocols in Molecular Biology
Richard Mortensen

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.

References

Jun 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·H te RieleA Berns
Jul 1, 1992·Molecular and Cellular Biology·K R ThomasM R Capecchi
Nov 1, 1991·Molecular and Cellular Biology·P HastyA Bradley
Dec 11, 1991·Proceedings of the National Academy of Sciences of the United States of America·B H KollerO Smithies
May 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·R L YenofskyJ W Pellow
Mar 8, 1990·Nature·H Zheng, J H Wilson
Jun 16, 1989·Science·M R Capecchi
Jun 1, 1987·Molecular and Cellular Biology·E A Wong, M R Capecchi
Dec 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·G R Martin
Jun 7, 1994·Proceedings of the National Academy of Sciences of the United States of America·S ChengR Higuchi

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Citations

Nov 23, 2012·Biotechnology Progress·Seshu TummalaAnthony Rossomando
Jan 27, 2009·Current Protocols in Human Genetics·Christopher Semsarian

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