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Cre Transgenic Mouse Lines

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 561))

Summary

With the development of the Cre-LoxP system, conditional gene targeting has rapidly become a powerful technology that facilitates the study of gene function. This advanced technique circumvents three major concerns sometimes levelled against conventional transgenic and gene-targeting approaches. First of all, gene ablation may exert its effect in multiple cell and tissue types, creating a complex phenotype in which it is difficult to distinguish direct function in a particular tissue from secondary effects resulting from altered gene function in other tissues. Secondly, a gene deletion expressed in the germ line may cause embryonic lethality, thereby precluding analysis of gene function in the adult tissues. Thirdly, the transgenic approach represents a somewhat surreal over-expression of a given protein often causing spurious phenotypes. The generation of conditional knockout mice is a multiple-step process, which involves mating the flox mutant mouse line (essential exon/s of the gene of interest are flanked by two LoxP sites) and the Cre-expressing mouse line. Over the past few years many inducible and/or tissue-specific Cre mouse lines have been developed. This chapter will give a brief review of the generation of Cre-expressing mouse lines and will discuss the strategy of using these Cre lines. In addition, information regarding established Cre-expressing mouse lines will be provided.

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References

  1. Kühn, R., Schwenk, F., Aguet, M., and Rajewsky, K. (1995). Inducible gene targeting in mice. Science, 269, 1427–1428

    Article  PubMed  Google Scholar 

  2. Feil, R., Brocard, J., Mascrez, B., LeMeur, M., Metzger, D., and Chambon, P. (1996). Ligand-activated site-specific recombination in mice. Proc. Natl. Acad. Sci. U.S.A., 93, 10887–10890

    Article  PubMed  CAS  Google Scholar 

  3. Soriano, P. (1999). Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet., 21, 70–71

    Article  PubMed  CAS  Google Scholar 

  4. Guy, J., Hendrich, B., Holmes, M., Martin, J.E., and Bird, A. (2001). A mouse Mecp2-null mutation causes neurological symptoms that mimic Rett syndrome. Nat Genet., 27(3), 322–326

    Article  PubMed  CAS  Google Scholar 

  5. Imayoshi, I., Ohtsuka, T., Metzger, D., Chambon, P., and Kegeyama, R. (2006). Temporal regulation of Cre recombinase activity in neural stem cells. Genesis, 44(5), 233–238

    Article  PubMed  CAS  Google Scholar 

  6. Casanova, E., Fehsenfeld, S., Mantamadiotis, T., Lemberger, T., Greiner, E., Stewart, A.F., and Schütz, G. (2001). A CamKIIalpha iCre BAC allows brain-specific gene inactivation. Genesis, 31(1), 37–42

    Article  PubMed  CAS  Google Scholar 

  7. Casanova, E., Fehsenfeld, S., Lemberger, T., Shimshek, D.R., Sprengel, R., and Mantamadiotis, T. (2002). ER-based double iCre fusion protein allows partial recombination in forebrain. Genesis, 34(3), 208–214

    Article  PubMed  CAS  Google Scholar 

  8. Fraser, M.M., Zhu, X., Kwon, C.H., Uhlmann, E.J., Gutmann, D.H., and Baker, S.J.(2004). Pten loss causes hypertrophy and increased proliferation of astrocytes in vivo. Cancer Res., 64(21), 7773–7779

    Article  PubMed  CAS  Google Scholar 

  9. Barski, J.J., Dethleffsen, K., and Meyer, M. (2000). Cre recombinase expression in cerebellar Purkinje cells. Genesis, 28(3-4), 93–98

    Article  PubMed  CAS  Google Scholar 

  10. Agah, R., Frenkel, P.A., French, B.A., Michael, L.H., Overbeek, P.A., and Schneider, M.D. (1997). Gene recombination in postmitotic cells. Targeted expression of Cre recombinase provokes cardiac-restricted, site-specific rearrangement in adult ventricular muscle in vivo. J. Clin. Invest., 100(1), 169–179

    Article  PubMed  CAS  Google Scholar 

  11. Sohal, D.S., Nghiem, M., Crackower, M.A., Witt, S.A., Kimball, T.R., Tymitz, K.M., Penninger, J.M., and Molkentin, J.D. (2001). Temporally regulated and tissue-specific gene manipulations in the adult and embryonic heart using a tamoxifen-inducible Cre protein. Circ Res, 89, 20–25

    Article  PubMed  CAS  Google Scholar 

  12. Chen, J., Kubalak, S.W., Minamisawa, S., Price, R.L., Becker, K.D., Hickey, R., Ross, J. Jr., and Chien, K.R. (1998). Selective requirement of myosin light chain 2v in embryonic heart function. J. Biol. Chem., 273(2), 1252–1256

    Article  PubMed  CAS  Google Scholar 

  13. Gerald, W.M.B., Haspel, J.A., Smith, X.C.L., Wiener, H.H., and Burden, S.J. (2000). Selective expression of Cre recombinase in skeletal muscle fibers. Genesis, 26, 165–166

    Article  Google Scholar 

  14. Xin, H.B., Deng, K.Y., Rishniw, M., Ji, G., and Kotlikoff, M.I. (2002). Smooth muscle expression of Cre recombinase and eGFP in transgenic mice. Physiol. Genomics, 10(3), 211–215

    PubMed  CAS  Google Scholar 

  15. Kellendonk, C., Opherk, C., Anlag, K., Schütz, G., and Tronche, F. (2000). Hepatocyte-specific expression of Cre recombinase. Genesis, 26(2), 151–153

    Article  PubMed  CAS  Google Scholar 

  16. Gannon, M., Shiota, C., Postic, C., Wright, C.V.E., and Magnuson, M. (2000). Analysis of the Cre-mediated recombination driven by rat insulin promoter in embryonic and adult mouse pancreas. Genesis, 26, 139–141

    Article  PubMed  CAS  Google Scholar 

  17. Maddison, L.A., Nahm, H., DeMayo, F., and Greenberg, N.M. (2000). Prostate specific expression of Cre recombinase in transgenic mice. Genesis, 26, 154–156

    Article  PubMed  CAS  Google Scholar 

  18. Leheste, J.R., Melsen, F., Wellner, M., Jansen, P., Schlichting, U., Renner-Müller, I., Andreassen, T.T., Wolf, E., Bachmann, S., nykjaer, A., and Willnow, T.E. (2003). Hypocalcemia and osteopathy in mice with kidney-specific megalin gene defect. FASEB J., 17(2), 247–249

    PubMed  CAS  Google Scholar 

  19. Marquardt, T., Ashery-Padan, R., Andrejewski, N., Scardigli, R., Guillemot, F., and Gruss, P. (2001). Pax6 is required for the multipotent state of retinal progenitor cells. Cell, 105, 43–55

    Article  PubMed  CAS  Google Scholar 

  20. Gustafsson , E., Brakebusch, C., Hietanen, K., and Fassler, R. (2001). Tie-1-directed expression of Cre recombinase in endothelial cells of embryoid bodies and transgenic mice. J. Cell Sci., 114, 671–676

    PubMed  CAS  Google Scholar 

  21. Kisanuki, Y.Y., Hammer, R.E., Miyazaki, J., Williams, S.C., Richardson, J.A., and Yanagisawa, M. (2001). Tie2-Cre transgenic mice: a new model for endothelial cell-lineage analysis in vivo. Dev. Biol., 230(2), 230–242

    Article  PubMed  CAS  Google Scholar 

  22. Gu, H., Marth, J.D., Orban, P.C., Mossmann, H., and Rajewsky, K. (1994). Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. Science, 265(5168), 103–106

    Article  PubMed  CAS  Google Scholar 

  23. Takeda, K., Clausen, B.E., Kaisho, T., Tsujimura, T., Terada, N., Förster, I., and Akira, S. (1999). Enhanced Th1 activity and development of chronic enterocolitis in mice devoid of Stat3 in macrophages and neutrophils. Immunity, 10(1), 39–49

    Article  PubMed  CAS  Google Scholar 

  24. Sano, S., Itami, S., Takeda, K., Tarutani, M., Yamaguchi, Y., Miura, H., Yoshikawa, K., Akira, S., and Takeda, J. (1999). Keratinocyte-specific ablation of Stat3 exhibits impaired skin remodeling, but does not affect skin morphogenesis. EMBO J., 18(17), 4657–4668

    Article  PubMed  CAS  Google Scholar 

  25. Vasioukhin, V., Degenstein, L., Wise, B., and Fuchs, E. (1999). The magical touch: genome targeting in epidermal stem cells induced by tamoxifen application to mouse skin. Proc. Natl. Acad. Sci. U.S.A., 96(15), 8551–8556

    Article  PubMed  CAS  Google Scholar 

  26. Wagner, K.U., Wall, R.J., St-Onge, L., Gruss, P., Wynshaw-Boris, A., Garrett, L., Li, M., Furth, P.A., and Hennighausen, L. (1997). Cre-mediated gene deletion in the mammary gland. Nucleic Acids Res., 25(21), 4323–4330

    Article  PubMed  CAS  Google Scholar 

  27. Ovchinnikov, D.A., Deng, J.M., Ogunrinu, G., and Behringer, R.R. (2000). Col2a1-directed expression of Cre recombinase in differentiating chondrocytes in transgenic mice. Genesis, 26, 145–146

    Article  PubMed  CAS  Google Scholar 

  28. Castro, C.H., Stains, J.P., Sheikh, S.,Szejnfeld, V.L., Willecke, K., Theis, M., and Civitelli, R. (2003). Development of mice with osteoblast-specific connexin43 gene deletion. Cell Commun. Adhes., 10(4-6), 445–450

    Article  PubMed  CAS  Google Scholar 

  29. Barlow, C., Schroeder, M., Lekstrom-Himes, J., Kylefjord, H., Deng, C.X., Wynshaw-Boris, A., Spiegelman, B.M., and Xanthopoulos, K.G. (1997). Targeted expression of Cre recombinase to adipose tissue of transgenic mice directs adipose-specific excision of loxP-flanked gene segments. Nucleic Acids Res., 25(12), 2543–2545

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Xin Wang .

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Wang, X. (2009). Cre Transgenic Mouse Lines. In: Cartwright, E. (eds) Transgenesis Techniques. Methods in Molecular Biology, vol 561. Humana Press. https://doi.org/10.1007/978-1-60327-019-9_17

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  • DOI: https://doi.org/10.1007/978-1-60327-019-9_17

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-018-2

  • Online ISBN: 978-1-60327-019-9

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