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Determination of Gene Function by Homologous Recombination Using Embryonic Stem Cells and Knockout Mice

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Genomics Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 175))

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Abstract

A decade ago the first alteration of a gene using embryonic stem (ES) cells was performed in the mouse (1). Since then ES cells have become a powerful tool to generate mutant mice for the analysis of gene function. These mutant mice (also called knockout mice) have provided new insights into normal and pathologic development to improve our knowledge and open new avenues into diagnostic and therapeutic fields.

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References

  1. Schwartzberg, P. L., Goff, S. P., and Robertson, E. J. (1989) Germ-line transmission of a c-abl mutation produced by targeted gene disruption in ES cells. Science 246, 799–803.

    Article  PubMed  CAS  Google Scholar 

  2. Capecchi, M. R. (1989) Altering the mouse genome by homologous recombination. Science 244, 1288–1292.

    Article  PubMed  CAS  Google Scholar 

  3. Hogan, B., Beddington, R., Constantini, F., and Lacy, E., eds. (1994) In vitro culture. In: Manipulating the Mouse Embryo: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp. 389–397.

    Google Scholar 

  4. Olson, E. N., Arnold, H. H., Rigby, P. W. J., and Wold, B. J. (1996) Know your neighbors: three phenotypes in Null mutants of myogenic bHLH gene MRF4. Cell 85, 1–4.

    Article  PubMed  CAS  Google Scholar 

  5. Sauer, B. (1993) Manipulation oftransgenes by site-specific recombination: use of Cre recombinase, in Methods in Enzymology (Wasserman, P. M. and DePamphilis, M. L., eds.), Academic, New York, pp. 890–900.

    Google Scholar 

  6. Hirota, H., Chen, J., Betz, U. A. K., Rajewsky, K., Gu, Y., Ross Jr., J., Muller, W., and Chien, K., R. (1999). Loss of a gp130 cardiac muscle cell survival pathway is a critical event in the onset of heart failure during biomechanical stress. Cell 97, 189–198.

    Article  PubMed  CAS  Google Scholar 

  7. Deng, C. and Capecchi, M. R. (1992) Reexamination of gene targeting frequency as a function of the extent of homology between the targeting vector and the target locus. Mol. Cell. Biol. 12, 3365–3371.

    PubMed  CAS  Google Scholar 

  8. Riele, H. T., Maandag, E. R., and Berns, A. (1992) High efficiency gene targeting in embryonic stem cells through homologous recombination with isogenic DNA constructs. Proc. Natl. Acad. Sci. USA 89, 5128–5132.

    Article  Google Scholar 

  9. Mansour, S. L., Thomas, K. R., and Capecchi, M. R. (1988) Disruption of the protoncogene ins-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature 336, 348–352.

    Article  PubMed  CAS  Google Scholar 

  10. Robertson, E. J. (1987) Embryo-derived stem cell lines, in Teratocarcinomas and Embryonic Stem Cells. A Practical Approach (Robertson, E. J., ed.), IRL, Oxford, pp. 71–112.

    Google Scholar 

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© 2001 Humana Press Inc., Totowa, NJ

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Mansouri, A. (2001). Determination of Gene Function by Homologous Recombination Using Embryonic Stem Cells and Knockout Mice. In: Starkey, M.P., Elaswarapu, R. (eds) Genomics Protocols. Methods in Molecular Biology™, vol 175. Humana Press. https://doi.org/10.1385/1-59259-235-X:397

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  • DOI: https://doi.org/10.1385/1-59259-235-X:397

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-774-8

  • Online ISBN: 978-1-59259-235-7

  • eBook Packages: Springer Protocols

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