Generation of transgenic frogs.
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.
Loeber
Jana
J
Department of Developmental Biochemistry, University of Goettingen, Goettingen, Germany.
Pan
Fong Cheng
FC
Pieler
Tomas
T
D000818:Animals; D030801:Animals, Genetically Modified; D015252:Deoxyribonucleases, Type II Site-Specific / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods*; D018014:Gene Transfer Techniques; D018095:Germ-Line Mutation; D008845:Microinjections / Q000379:methods; D009865:Oocytes / Q000166:cytology / Q000502:physiology; D011401:Promoter Regions, Genetic; D029701:Saccharomyces cerevisiae Proteins / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution; D019076:Transgenes / Q000502:physiology*; D014982:Xenopus laevis / Q000196:embryology / Q000235:genetics*
D029701:Saccharomyces cerevisiae Proteins; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific