title: An inducible transgene expression system for zebrafish and chick.
abstract: We have generated an inducible system to control the timing of transgene expression in zebrafish and chick. An estrogen receptor variant (ERT2) fused to the GAL4 transcriptional activator rapidly and robustly activates transcription within 3 hours of treatment with the drug 4-hydroxy-tamoxifen (4-OHT) in tissue culture and transgenic zebrafish. We have generated a broadly expressed inducible ERT2-GAL4 zebrafish line using the ubiquitin (ubi) enhancer. In addition, use of ERT2-GAL4 in conjunction with tissue-specific enhancers enables the control of transgene expression in both space and time. This spatial restriction and the ability to sustain forced expression are important advantages over the currently used heat-shock promoters. Moreover, in contrast to currently available TET and LexA systems, which require separate constructs with their own unique recognition sequences, ERT2-GAL4 is compatible with the growing stock of UAS lines being generated in the community. We also applied the same inducible system to the chick embryo and find that it is fully functional, suggesting that this strategy is generally applicable.
authors: MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK. sg13@sanger.ac.uk
authors: Sebastian S
authors: SS
authors: Gerety
authors: Marie A
authors: MA
authors: Breau
authors: Noriaki
authors: N
authors: Sasai
authors: Qiling
authors: Q
authors: Xu
authors: James
authors: J
authors: Briscoe
authors: David G
authors: DG
authors: Wilkinson
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002642:Chick Embryo; D003433:Crosses, Genetic; D015509:Developmental Biology / Q000379:methods*; D018507:Gene Expression Regulation, Developmental; D057809:HEK293 Cells; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D010641:Phenotype; D011960:Receptors, Estrogen / Q000235:genetics* / Q000378:metabolism; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology; D019076:Transgenes; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000378:metabolism
chemical_list: D011960:Receptors, Estrogen; D029961:Zebrafish Proteins; D013629:Tamoxifen; C016601:afimoxifene
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