Refined spatial manipulation of neuronal function by combinatorial restriction of transgene expression

Neuron. 2006 Nov 9;52(3):425-36. doi: 10.1016/j.neuron.2006.08.028.

Abstract

Selective genetic manipulation of neuronal function in vivo requires techniques for targeting gene expression to specific cells. Existing systems accomplish this using the promoters of endogenous genes to drive expression of transgenes directly in cells of interest or, in "binary" systems, to drive expression of a transcription factor or recombinase that subsequently activates the expression of other transgenes. All such techniques are constrained by the limited specificity of the available promoters. We introduce here a combinatorial system in which the DNA-binding (DBD) and transcription-activation (AD) domains of a transcription factor are independently targeted using two different promoters. The domains heterodimerize to become transcriptionally competent and thus drive transgene expression only at the intersection of the expression patterns of the two promoters. We use this system to dissect a neuronal network in Drosophila by selectively targeting expression of the cell death gene reaper to subsets of neurons within the network.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Intramural

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Cells, Cultured
  • DNA-Binding Proteins / physiology
  • Drosophila
  • Drosophila Proteins / metabolism
  • Gene Expression / physiology*
  • Gene Expression Regulation*
  • Genetic Vectors* / genetics
  • Green Fluorescent Proteins / metabolism
  • Immunohistochemistry / methods
  • Neurons / classification
  • Neurons / physiology*
  • Transcription Factors / physiology
  • Transfection / methods
  • Transgenes*
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • DNA-Binding Proteins
  • Drosophila Proteins
  • Transcription Factors
  • rpr protein, Drosophila
  • Green Fluorescent Proteins
  • beta-Galactosidase