title: Spatial and temporal control of transgene expression in zebrafish.
abstract: Transgenic zebrafish research has provided valuable insights into gene functions and cell behaviors directing vertebrate development, physiology, and disease models. Most approaches use constitutive transgene expression and therefore do not provide control over the timing or levels of transgene induction. We describe an inducible gene expression system that uses new tissue-specific zebrafish transgenic lines that express the Gal4 transcription factor fused to the estrogen-binding domain of the human estrogen receptor. We show these Gal4-ERT driver lines confer rapid, tissue-specific induction of UAS-controlled transgenes following tamoxifen exposure in both embryos and adult fish. We demonstrate how this technology can be used to define developmental windows of gene function by spatiotemporal-controlled expression of constitutively active Notch1 in embryos. Given the array of existing UAS lines, the modular nature of this system will enable many previously intractable zebrafish experiments.
authors.affiliation: Institute of Molecular Biology, University of Oregon, Eugene, Oregon, United States of America; Department of Biology, University of Oregon, Eugene, Oregon, United States of America.
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Akerberg
authors.affiliation: Institute of Molecular Biology, University of Oregon, Eugene, Oregon, United States of America.
authors.forename: Scott
authors.initials: S
authors.lastname: Stewart
authors.affiliation: Institute of Molecular Biology, University of Oregon, Eugene, Oregon, United States of America; Department of Biology, University of Oregon, Eugene, Oregon, United States of America.
authors.forename: Kryn
authors.initials: K
authors.lastname: Stankunas
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004268:DNA-Binding Proteins / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D009928:Organ Specificity; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D011401:Promoter Regions, Genetic; D011817:Rabbits; D051881:Receptor, Notch1 / Q000096:biosynthesis / Q000235:genetics; D011960:Receptors, Estrogen / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics*; D013629:Tamoxifen / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects*; D019076:Transgenes; D029867:Xenopus Proteins / Q000235:genetics; D014982:Xenopus laevis / Q000235:genetics; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism; D029961:Zebrafish Proteins / Q000235:genetics; D055544:beta-Globins / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; C542436:Gal4 protein, zebrafish; D020648:Peptide Elongation Factor 1; D051881:Receptor, Notch1; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D029867:Xenopus Proteins; D029961:Zebrafish Proteins; D055544:beta-Globins; C485184:enhanced green fluorescent protein; D013629:Tamoxifen; D049452:Green Fluorescent Proteins