Spatiotemporal gene expression targeting with the TARGET and gene-switch systems in Drosophila

Sci STKE. 2004 Feb 12;2004(220):pl6. doi: 10.1126/stke.2202004pl6.

Abstract

Targeted gene expression has become a standard technique for the study of biological questions in Drosophila. Until recently, transgene expression could be targeted in the dimension of either time or space, but not both. Several new systems have recently been developed to direct transgene expression simultaneously in both time and space. We describe here two such systems that we developed in our laboratory. The first system provides a general method for temporal and regional gene expression targeting (TARGET) with the conventional GAL4-upstream activator sequence (UAS) system and a temperature-sensitive GAL80 molecule, which represses GAL4 transcriptional activity at permissive temperatures. The second system, termed Gene-Switch, is based on a GAL4-progesterone receptor chimera that is hormone-inducible. We have used both systems for simultaneous spatial and temporal rescue of memory dysfunction in the rutabaga (rut) memory mutant of Drosophila. In this protocol, we provide guidelines for the use of these two novel systems, which should have general utility in studying Drosophila biology and in using the fly as a model for human disease.

MeSH terms

  • 5' Flanking Region / genetics
  • Animals
  • Animals, Genetically Modified
  • Drosophila melanogaster / genetics*
  • Gene Expression Regulation / genetics*
  • Gene Expression Regulation, Developmental / genetics
  • Gene Targeting / methods*
  • Genes, Insect / genetics
  • Genes, Switch / drug effects
  • Genes, Switch / genetics
  • Genetic Vectors / genetics
  • Larva / genetics
  • Mifepristone / pharmacology
  • Muscles / chemistry
  • Muscles / metabolism
  • Myosin Heavy Chains / genetics
  • Phenotype
  • Promoter Regions, Genetic
  • Regulatory Sequences, Nucleic Acid / genetics
  • Repressor Proteins / genetics
  • Sucrose / pharmacology
  • Transcriptional Activation / genetics
  • Transgenes / genetics

Substances

  • Repressor Proteins
  • Mifepristone
  • Sucrose
  • Myosin Heavy Chains