Generation of Transgenic Frogs

  • Jana Loeber
  • Fong Cheng Pan
  • Tomas Pieler
Part of the Methods in Molecular Biology book series (MIMB, volume 561)


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.

Key words

Xenopus laevis Transgenesis Meganuclease I-SceI Germline transmission Tissue-specific promoter 



This work was supported by funds from the Deutsche Forschungsgemeinschaft to T.P.


  1. 1.
    Kroll, K.L. and Amaya, E. (1996) Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation. Development, 122, 3173–3183.PubMedGoogle Scholar
  2. 2.
    Allen, B.G. and Weeks, D.L. (2005) Transgenic Xenopus laevis embryos can be generated using phiC31 integrase. Nat Methods, 2, 975–979.PubMedCrossRefGoogle Scholar
  3. 3.
    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.PubMedCrossRefGoogle Scholar
  4. 4.
    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.PubMedCrossRefGoogle Scholar
  5. 5.
    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.PubMedCrossRefGoogle Scholar
  6. 6.
    Ogino, H., McConnell, W.B. and Grainger, R.M. (2006) High-throughput transgenesis in Xenopus using I-SceI meganuclease. Nat Protoc, 1, 1703–1710.PubMedCrossRefGoogle Scholar
  7. 7.
    Beck, C.W. and Slack, J.M. (1999) Gut specific expression using mammalian promoters in transgenic Xenopus laevis. Mech Dev, 88, 221–227.PubMedCrossRefGoogle Scholar
  8. 8.
    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.PubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Jana Loeber
    • 1
  • Fong Cheng Pan
    • 2
    • 3
  • Tomas Pieler
    • 1
  1. 1.Department of Developmental BiochemistryUniversity of GoettingenGoettingenGermany
  2. 2.Vanderbilt University Program in Developmental Biology3144A MRBIII Vanderbilt University Medical CenterNashvilleTN
  3. 3.Department of Cell and Biology3144A MRBIII Vanderbilt University Medical CenterNashvilleTN

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