Transcriptional regulation using the Q system in transgenic zebrafish
- Ghosh, A., Halpern, M.E.
- Methods in cell biology 135: 205-18 (Chapter)
- Registered Authors
- Halpern, Marnie E.
- Bipartite system, Gal4, Transcriptional regulation, Transgenesis, qalocus
- MeSH Terms
- Animals, Genetically Modified/genetics
- DNA Transposable Elements/genetics*
- Enhancer Elements, Genetic/genetics*
- Gene Expression Regulation, Developmental
- Green Fluorescent Proteins/genetics
- Neurospora crassa/genetics
- Transcription Factors/genetics*
- Transcriptional Activation/genetics*
- 27443927 Full text @ Meth. Cell. Biol.
Ghosh, A., Halpern, M.E. (2016) Transcriptional regulation using the Q system in transgenic zebrafish. Methods in cell biology. 135:205-18.
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.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes