title: Site-specific transgenesis in Xenopus.
abstract: Transgenesis is an essential, powerful tool for investigating gene function and the activities of enhancers, promoters, and transcription factors in the chromatin environment. In Xenopus, current methods generate germ-line transgenics by random insertion, often resulting in mosaicism, position-dependent variations in expression, and lab-to-lab differences in efficiency. We have developed and tested a Xenopus FLP-FRT recombinase-mediated transgenesis (X-FRMT) method. We demonstrate transgenesis of Xenopus laevis by FLP-catalyzed recombination of donor plasmid cassettes into F(1) tadpoles with host cassette transgenes. X-FRMT provides a new method for generating transgenic Xenopus. Once Xenopus lines harboring single host cassettes are generated, X-FRMT should allow for the targeting of transgenes to well-characterized integration site(s), requiring no more special reagents or training than that already common to most Xenopus labs.
authors.affiliation: Department of Ophthalmology, The Center for Vision Research and the SUNY Eye Institute, Upstate Medical University, Syracuse, NY 13210, USA. zuberm@upstate.edu
authors.forename: Michael E
authors.initials: ME
authors.lastname: Zuber
authors.forename: Heather S
authors.initials: HS
authors.lastname: Nihart
authors.forename: Xinming
authors.initials: X
authors.lastname: Zhuo
authors.forename: Sudha
authors.initials: S
authors.lastname: Babu
authors.forename: Barry E
authors.initials: BE
authors.lastname: Knox
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D005260:Female; D023061:Gene Order; D018390:Gene Targeting / Q000379:methods*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D008297:Male; D011995:Recombination, Genetic; D019076:Transgenes; D014982:Xenopus laevis / Q000235:genetics*
chemical_list: D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase