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Generating mice with targeted mutations

Nature Medicine volume 7, pages 1086–1090 (2001)Cite this article

Mutational analysis is one of the most informative approaches available for the study of complex biological processes. It has been particularly successful in the analysis of the biology of bacteria, yeast, the nematode worm Caenorhabditis elegans and the fruit fly Drosophila melanogaster. Extension of this approach to the mouse, though informative, was far less successful relative to what has been achieved with these simpler model organisms. This is because it is not numerically practical in mice to use random mutagenesis to isolate mutations that affect a specified biological process of interest. Nonetheless, biological phenomena such as a sophisticated immune response, cancer, vascular disease or higher-order cognitive function, to mention just a few, must be analyzed in organisms that show such phenomena, and for this reason geneticists and other researchers have turned to the mouse. Gene targeting, the means for creating mice with designed mutations in almost any gene32, was developed as an alternative to the impractical use of random mutagenesis for pursuing genetic analysis in the mouse. Now gene targeting has advanced the genomic manipulations possible in mice to a level that can be matched only in far simpler organisms such as bacteria and yeast.

The development of gene targeting in mice required the solution to two problems: How to produce a specific mutation in a chosen gene in cultured mammalian cells, and how to transfer this mutation to the mouse germ line. Oliver Smithies' laboratory and mine worked independently on solutions to the first problem. Martin Evans' laboratory provided the basis for a solution to the second problem.

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Figure 1: Regenera-tion of a functional neor by gene targeting.
Figure 2: Disruption of Hprt by gene targeting.
Figure 3: The positive–negative selection procedure used to enrich for ES cells containing a targeted disruption of gene X.
Figure 4: Generation of mouse germline chimeras from ES cells containing a targeted mutation.

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  1. Howard Hughes Medical Institute Department of Human Genetics University of Utah School of Medicine, Salt Lake City, Utah, USA

    Mario R. Capecchi

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Capecchi, M. Generating mice with targeted mutations. Nat Med 7, 1086–1090 (2001). https://doi.org/10.1038/nm1001-1086

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