Transactivation from Gal4-VP16 transgenic insertions for tissue-specific cell labeling and ablation in zebrafish

Jon M. Davison, Courtney M. Akitake, Mary G. Goll, Jerry M. Rhee, Nathan Gosse, Herwig Baier, Marnie E. Halpern, Steven D. Leach, Michael J. Parsons

Research output: Contribution to journalArticlepeer-review

Abstract

Prior studies with transgenic zebrafish confirmed the functionality of the transcription factor Gal4 to drive expression of other genes under the regulation of upstream activator sequences (UAS). However, widespread application of this powerful binary system has been limited, in part, by relatively inefficient techniques for establishing transgenic zebrafish and by the inadequacy of Gal4 to effect high levels of expression from UAS-regulated genes. We have used the Tol2 transposition system to distribute a self-reporting gene/enhancer trap vector efficiently throughout the zebrafish genome. The vector uses the potent, hybrid transcription factor Gal4-VP16 to activate expression from a UAS:eGFP reporter cassette. In a pilot screen, stable transgenic lines were established that express eGFP in reproducible patterns encompassing a wide variety of tissues, including the brain, spinal cord, retina, notochord, cranial skeleton and muscle, and can transactivate other UAS-regulated genes. We demonstrate the utility of this approach to track Gal4-VP16 expressing migratory cells in UAS:Kaede transgenic fish, and to induce tissue-specific cell death using a bacterial nitroreductase gene under UAS control. The Tol2-mediated gene/enhancer trapping system together with UAS transgenic lines provides valuable tools for regulated gene expression and for targeted labeling and ablation of specific cell types and tissues during early zebrafish development.

Original languageEnglish (US)
Pages (from-to)811-824
Number of pages14
JournalDevelopmental biology
Volume304
Issue number2
DOIs
StatePublished - Apr 15 2007

Keywords

  • Cell tracing
  • Enhancer trap
  • Gene trap
  • Nitroreductase
  • Photoconversion
  • Tol2 transposon

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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