Summary
The possibility of generating transgenic animals is of obvious advantage for the analysis of gene function in development and disease. One of the established vertebrate model systems in developmental biology is the amphibian Xenopus laevis. Different techniques have been successfully applied to create Xenopus transgenics; in this chapter, the so-called meganuclease method is described. This technique is not only technically simple, but also comparably efficient and applicable to both Xenopus laevis and Xenopus tropicalis. The commercially available endonuclease I-SceI (meganuclease) mediates the integration of foreign DNA into the frog genome after coinjection into fertilized eggs. Tissue-specific gene expression, as well as germline transmission, has been observed.
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References
Kroll, K.L. and Amaya, E. (1996) Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation. Development, 122, 3173–3183.
Allen, B.G. and Weeks, D.L. (2005) Transgenic Xenopus laevis embryos can be generated using phiC31 integrase. Nat Methods, 2, 975–979.
Thermes, V., Grabher, C., Ristoratore, F., Bourrat, F., Choulika, A., Wittbrodt, J. and Joly, J.S. (2002) I-SceI meganuclease mediates highly efficient transgenesis in fish. Mech Dev, 118, 91–98.
Pan, F.C., Chen, Y., Loeber, J., Henningfeld, K. and Pieler, T. (2006) I-SceI meganuclease-mediated transgenesis in Xenopus. Dev Dyn, 235, 247–252.
Ogino, H., McConnell, W.B. and Grainger, R.M. (2006) Highly efficient transgenesis in Xenopus tropicalis using I-SceI meganuclease. Mech Dev, 123, 103–113.
Ogino, H., McConnell, W.B. and Grainger, R.M. (2006) High-throughput transgenesis in Xenopus using I-SceI meganuclease. Nat Protoc, 1, 1703–1710.
Beck, C.W. and Slack, J.M. (1999) Gut specific expression using mammalian promoters in transgenic Xenopus laevis. Mech Dev, 88, 221–227.
Offield, M.F., Hirsch, N. and Grainger, R.M. (2000) The development of Xenopus tropicalis transgenic lines and their use in studying lens developmental timing in living embryos. Development, 127, 1789–1797.
Acknowledgments
This work was supported by funds from the Deutsche Forschungsgemeinschaft to T.P.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Loeber, J., Pan, F.C., Pieler, T. (2009). Generation of Transgenic Frogs. In: Cartwright, E. (eds) Transgenesis Techniques. Methods in Molecular Biology, vol 561. Humana Press. https://doi.org/10.1007/978-1-60327-019-9_4
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DOI: https://doi.org/10.1007/978-1-60327-019-9_4
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