Use of the Gal4-UAS Technique for Targeted Gene Expression in the Zebrafish

Mech Dev. 1999 Feb;80(2):153-8. doi: 10.1016/s0925-4773(98)00209-3.

Abstract

The most common way to analyze the function of cloned genes in zebrafish is to misexpress the gene product or an altered variant of it by mRNA injection. However, mRNA injection has several disadvantages. The GAL4-UAS system for targeted gene expression allows one to overcome some of these disadvantages. To test the GAL4-UAS system in zebrafish, we generated two different kinds of stable transgenic lines, carrying activator and effector constructs, respectively. In the activator lines the gene for the yeast transcriptional activator GAL4 is under the control of a given promoter, while in the effectors the gene of interest is fused to the sequence of the DNA-binding motif of GAL4 (UAS). Crosses of animals from the activator and effector lines show that effector genes are transcribed with the spatial pattern of the activators. This work smoothes the way for a novel method of misexpression of gene products in zebrafish in order to analyze the function of genes in developmental processes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics
  • Adenovirus E1B Proteins / genetics
  • Animals
  • Animals, Genetically Modified
  • Cloning, Molecular
  • Crosses, Genetic
  • DNA-Binding Proteins
  • Embryo, Nonmammalian / metabolism
  • Enhancer Elements, Genetic
  • Fungal Proteins / genetics*
  • Gene Expression Regulation, Developmental*
  • Genes, Reporter*
  • Genes, myc
  • In Situ Hybridization
  • Membrane Proteins / biosynthesis
  • Membrane Proteins / genetics
  • Pilot Projects
  • Promoter Regions, Genetic*
  • Proto-Oncogene Proteins c-myc / biosynthesis
  • Receptor, Notch1
  • Receptors, Cell Surface*
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / physiology
  • Regulatory Sequences, Nucleic Acid*
  • Saccharomyces cerevisiae Proteins*
  • Simian virus 40 / genetics
  • Simplexvirus / enzymology
  • Simplexvirus / genetics
  • Thymidine Kinase / genetics
  • Transcription Factors / genetics*
  • Transcription, Genetic*
  • Transgenes
  • Viral Proteins / genetics
  • Zebrafish / embryology
  • Zebrafish / genetics*

Substances

  • Actins
  • Adenovirus E1B Proteins
  • DNA-Binding Proteins
  • Fungal Proteins
  • GAL4 protein, S cerevisiae
  • Membrane Proteins
  • Proto-Oncogene Proteins c-myc
  • Receptor, Notch1
  • Receptors, Cell Surface
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Viral Proteins
  • Thymidine Kinase