Transcriptional regulation using the Q system in transgenic zebrafish

A. Ghosh, Marnie Halpern

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationThe Zebrafish Genetics, Genomics, and Transcriptomics, 2016
PublisherAcademic Press Inc.
Pages205-218
Number of pages14
Volume135
ISBN (Print)9780128034743
DOIs
Publication statusPublished - 2016
Externally publishedYes

Publication series

NameMethods in Cell Biology
Volume135
ISSN (Print)0091679X

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Keywords

  • Bipartite system
  • Gal4
  • qalocus
  • Transcriptional regulation
  • Transgenesis

ASJC Scopus subject areas

  • Cell Biology

Cite this

Ghosh, A., & Halpern, M. (2016). Transcriptional regulation using the Q system in transgenic zebrafish. In The Zebrafish Genetics, Genomics, and Transcriptomics, 2016 (Vol. 135, pp. 205-218). (Methods in Cell Biology; Vol. 135). Academic Press Inc.. https://doi.org/10.1016/bs.mcb.2016.05.001