Protocol

Hedgehog Signaling Protocols

Volume 397 of the series Methods Inmolecular Biology™ pp 161-201

GAL4/UAS Targeted Gene Expression for Studying Drosophila Hedgehog Signaling

  • Denise BussonAffiliated withInstitut Jacques Monod (UMR7592-CNRS/Universités Paris VI et VII)
  • , Anne-Marie PretAffiliated withCentre de Génétique Moléculaire (UPR2167-CNRS associé à l’Université Paris VI), de la Terrasse Gif sur Yvette

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Abstract

The GAL4/upstream activating sequence (UAS) system is one of the most powerful tools for targeted gene expression. It is based on the properties of the yeast GAL4 transcription factor which activates transcription of its target genes by binding to UAS cis-regulatory sites. In Drosophila, the two components are carried in separate lines allowing for numerous combinatorial possibilities. The driver lines provide tissue-specific GAL4 expression and the responder lines carry the coding sequence for the gene of interest under the control of UAS sites. In this chapter, the basic GAL4/UAS system and its extensions, namely those allowing precise temporal control and reversible expression, are described. In addition, a list of GAL4 and UAS lines and schematic maps of GAL4 and UAS vectors useful in the study of Hedgehog (Hh) signaling is given. Finally, uses of the GAL4/UAS system to resolve some of the questions addressed in the study of the Hh pathway are presented.

Key Words

GAL4 UAS GAL80 FLP/FRT GAL4 driver lines UAS responder lines targeted gene expression spatio-temporal control reversible expression gain-of-function