title: Transcriptional regulation using the Q system in transgenic zebrafish.
abstract: Methods to label cell populations selectively or to modify their gene expression are critical tools in the study of developmental or physiological processes in vivo. A variety of approaches have been applied to the zebrafish model, capitalizing on Tol2 transposition to generate transgenic lines with high efficiency. Here we describe the adoption of the Q system of Neurospora crassa, which includes the QF transcription factor and the upstream activating sequence (QUAS) to which it binds. These components function as a bipartite regulatory system similar to that of yeast Gal4/UAS, producing robust expression in transient assays of zebrafish embryos injected with plasmids and in stable transgenic lines. An important advantage, however, is that QUAS-regulated transgenes appear far less susceptible to transcriptional silencing even after seven generations. This chapter describes some of the Q system reagents that have been developed for zebrafish, as well as the use of the QF transcription factor for isolation of tissue-specific driver lines from gene/enhancer trap screens. Additional strategies successfully implemented in invertebrate models, such as a truncated QF transcription factor (QF2) or the reassembly of a split QF, are also discussed. The provided information, and available Gateway-based vectors, should enable those working with the zebrafish model to implement the Q system with minimal effort or to use it in combination with Gal4, Cre, or other regulatory systems for further refinement of transcriptional control.
authors.affiliation: Carnegie Institution for Science, Baltimore, MD, United States.
authors.forename: A
authors.initials: A
authors.lastname: Ghosh
authors.affiliation: Carnegie Institution for Science, Baltimore, MD, United States.
authors.forename: M E
authors.initials: ME
authors.lastname: Halpern
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins / Q000235:genetics; D009492:Neurospora crassa / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*; D015533:Transcriptional Activation / Q000235:genetics*; D019076:Transgenes / Q000235:genetics; D015027:Zebrafish / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements; D014157:Transcription Factors; D049452:Green Fluorescent Proteins
keywords: Bipartite system; Gal4; Transcriptional regulation; Transgenesis; qalocus