Abstract
Integrins and cell adhesion molecules are required for cell adhesion and movement, as well as various biological functions such as development or immune responses. To elucidate their in vivo functions, mice lacking these molecules have been generated and various phenotypes analyzed. In the case of embryonic lethality or a requirement for spatiotemporal ablation of these genes, conditional gene-targeted mice have also been generated. Gene targeting is a powerful tool to study the function of proteins under physiological conditions and is a widely used method. Construction of targeting vectors and selection of homologous recombinant embryonic stem cells require delicate and elaborate technical procedures. In this chapter we describe the methods to generate conventional and conditional knockout mice.
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References
Hynes, R.O. (1992) Integrins: versatility, modulation, and signaling in cell adhesion. Cell 69, 11–25.
Hynes, R.O. (2002) Integrins: bidirectional, allosteric signaling machines. Cell 110, 673–687.
Frenette, P.S., and Wagner, D.D. (1997) Insights into selectin function from knockout mice. Thromb. Haemost. 78, 60–64.
Evans, M.J., and Kaufman, M.H. (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154–156.
Evans, M.J. (1989) Potential for genetic manipulation of mammals. Mol. Biol. Med. 6, 557–565.
Zijlstra, M., Li, E., Sajjadi, F., Subramani, S., and Jaenisch, R. (1989) Germ-line transmission of a disrupted beta 2-microglobulin gene produced by homologous recombination in embryonic stem cells. Nature 342, 435–438.
Yang, J.T., Rayburn, H., and Hynes, R.O. (1993) Embryonic mesodermal defects in alpha 5 integrin-deficient mice. Development 119, 1093–1105.
Fässler, R., and Meyer, M. (1995) Consequences of lack of beta 1 integrin gene expression in mice. Genes Dev. 9, 1896–1908.
Kühn, R., Schwenk, F., Aguet, M., and Rajewsky, K. (1995). Inducible gene targeting in mice. Science 269, 1427–1428.
Nagy A. (2000) Cre recombinase: The universal reagent for genome tailoring. Genesis 26, 99–109.
Branda, C.S., and Dymecki, S.M. (2004) Talking about a revolution: The impact of site-specific recombinases on genetic analyses in mice. Dev. Cell 6, 7–28.
Wang, X. (2009) Cre transgenic mouse lines. Methods Mol. Biol. 561, 265–273.
Acknowledgments
We thank C. Hidaka for excellent secretarial assistance, Y. Magota for technical assistance, and K. Kimura for figure making. This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology, the Strategic Inter-national Cooperative Program (Research Exchange Type), the Japan Science and Technology Agency (JST), and the Takeda Science Foundation; KANAE FOUNDATION FOR THE PROMO-TION OF MEDICAL SCIENCE; The Cell Science Research Foundation; THE ICHIRO KANEHARA FOUNDATION; Kato Memorial Bioscience Foundation; and THE UEHARA MEM-ORIAL FOUNDATION. The authors have no conflicting financial interests.
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Yamamoto, M., Takeda, K. (2011). A Method for the Generation of Conditional Gene-Targeted Mice. In: Shimaoka, M. (eds) Integrin and Cell Adhesion Molecules. Methods in Molecular Biology, vol 757. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-166-6_23
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DOI: https://doi.org/10.1007/978-1-61779-166-6_23
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Publisher Name: Humana Press, Totowa, NJ
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