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The Tol2-mediated Gal4-UAS method for gene and enhancer trapping in zebrafish.

Methods (San Diego, Calif.) | 2009

The Gal4-UAS system provides powerful tools to analyze the function of genes and cells in vivo and has been extensively employed in Drosophila. The usefulness of this approach relies on the P element-mediated Gal4 enhancer trapping, which can efficiently generate transgenic fly lines expressing Gal4 in specific cells. Similar approaches, however, had not been developed in vertebrate systems due to the lack of an efficient transgenesis method. We have been developing transposon techniques by using the madaka fish Tol2 element. Taking advantage of its ability to generate genome-wide insertions, we developed the Gal4 gene trap and enhancer trap methods in zebrafish that enabled us to create various transgenic fish expressing Gal4 in specific cells. The Gal4-expressing cells can be visualized and manipulated in vivo by crossing the transgenic Gal4 lines with transgenic lines carrying various reporter and effector genes downstream of UAS (upstream activating sequence). Thus, the Gal4 gene trap and enhancer trap methods together with UAS lines now make detailed analyses of genes and cells in zebrafish feasible. Here, we describe the protocols to perform Gal4 gene trap and enhancer trap screens in zebrafish and their application to the studies of vertebrate neural circuits.

Pubmed ID: 19835787 RIS Download

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Associated grants

  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM069382-02
  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM069382-03
  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM069382
  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM069382-01
  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM069382-04

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Zebrafish Information Network (ZFIN) (tool)

RRID:SCR_002560

Model organism database that serves as central repository and web-based resource for zebrafish genetic, genomic, phenotypic and developmental data. Data represented are derived from three primary sources: curation of zebrafish publications, individual research laboratories and collaborations with bioinformatics organizations. Data formats include text, images and graphical representations.Serves as primary community database resource for laboratory use of zebrafish. Developed and supports integrated zebrafish genetic, genomic, developmental and physiological information and link this information extensively to corresponding data in other model organism and human databases.

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zn-12 (antibody)

RRID:AB_10013761

This monoclonal targets zn-12

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