title: A targeted gene expression system using the tryptophan repressor in zebrafish shows no silencing in subsequent generations.
abstract: The ability to visualize and manipulate cell fate and gene expression in specific cell populations has made gene expression systems valuable tools in developmental biology studies. Here, we describe a new system that uses the E. coli tryptophan repressor and its upstream activation sequence (TrpR/tUAS) to drive gene expression in stable zebrafish transgenic lines and in mammalian cells. We show that TrpR/tUAS transgenes are not silenced in subsequent generations of zebrafish, which is a major improvement over some of the existing systems, such as Gal4/gUAS and the Q-system. TrpR transcriptional activity can be tuned by mutations in its DNA-binding domain, or silenced by Gal80 when fused to the Gal4 activation domain. In cases in which more than one cell population needs to be manipulated, TrpR/tUAS can be used in combination with other, existing systems.
authors.affiliation: Department of Biological Structure, University of Washington, Seattle, WA 98195-5502, USA.
authors.forename: Arminda
authors.initials: A
authors.lastname: Suli
authors.forename: Ali D
authors.initials: AD
authors.lastname: Guler
authors.forename: David W
authors.initials: DW
authors.lastname: Raible
authors.forename: David
authors.initials: D
authors.lastname: Kimelman
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001426:Bacterial Proteins / Q000235:genetics*; D020868:Gene Silencing / Q000502:physiology*; D012097:Repressor Proteins / Q000235:genetics*; D015027:Zebrafish; D029968:Escherichia coli Proteins
chemical_list: D001426:Bacterial Proteins; D012097:Repressor Proteins; C027190:TRPR protein, E coli; D029968:Escherichia coli Proteins
keywords: Gal4/UAS; Gene expression system; Silencing; Tryptophan repressor; Zebrafish