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Gene targeting in mice: functional analysis of the mammalian genome for the twenty-first century

Nature Reviews Genetics volume 6, pages 507–512 (2005)Cite this article

Abstract

Gene targeting in mouse embryonic stem cells has become the 'gold standard' for determining gene function in mammals. Since its inception, this technology has revolutionized the study of mammalian biology and human medicine. Here I provide a personal account of the work that led to the generation of gene targeting which now lies at the centre of functional genomic analysis.

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Figure 1: Generation of an integrated, head-to-tail concatemer of multiple copies of a plasmid following its injection into nuclei of cultured mammalian cells.
Figure 2: Generation of a functional neomycin resistance gene from two defective genes by gene targeting.
Figure 3: Disruption of the endogenous hypoxanthine phosphoribosyl transferase gene by gene targeting in embryonic stem cells.
Figure 4: Positive–negative selection.
Figure 5: Generation of mouse germline chimaeras from embryonic stem cells that contain the desired targeted mutation.

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Acknowledgements

I would like to thank all those who have worked in my laboratory.

Author information

Authors and Affiliations

  1. Department of Human Genetics, Howard Hughes Medical Institute, University of Utah School of Medicine, 15 North 2030 East, Room 5100, Salt Lake City, 84112-5331, Utah, USA

    Mario R. Capecchi

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DATABASES

Entrez Gene

Hprt1

HSV-tk

FURTHER INFORMATION

Mario Capecchi's homepage

Glossary

ALLELIC SERIES

A series of alleles that can be present at the same locus, which produces graded phenotypes.

HYPOMORPHIC MUTATION

A mutation of which the phenotypic effects are less severe relative to a null mutation in the same gene.

PLURIPOTENT

When referring to stem cells, having the capacity to contribute to the formation of all cell types, such as embryonic stem cells.

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Capecchi, M. Gene targeting in mice: functional analysis of the mammalian genome for the twenty-first century. Nat Rev Genet 6, 507–512 (2005). https://doi.org/10.1038/nrg1619

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