The Tol2-mediated Gal4-UAS method for gene and enhancer trapping in zebrafish
- Asakawa, K., and Kawakami, K.
- Methods (San Diego, Calif.) 49(3): 275-281 (Journal)
- Registered Authors
- Kawakami, Koichi
- Gal4FF, Target gene expression, Transposition, Tetanus toxin light chain, Neural circuits
- MeSH Terms
- Animals, Genetically Modified
- Enhancer Elements, Genetic*
- Fungal Proteins/genetics
- Genes, Reporter
- Genetic Engineering/methods*
- Mutagenesis, Insertional/methods
- Zebrafish Proteins/genetics
- 19835787 Full text @ Methods
Asakawa, K., and Kawakami, K. (2009) The Tol2-mediated Gal4-UAS method for gene and enhancer trapping in zebrafish. Methods (San Diego, Calif.). 49(3):275-281.
The Gal4-UAS system provides powerful tools to analyze the function of genes and cells in vivo and has been extensively employed in Drosophila. The usefulness of this approach relies on the P element-mediated Gal4 enhancer trapping, which can efficiently generate transgenic fly lines expressing Gal4 in specific cells. Similar approaches, however, had not been developed in vertebrate systems due to the lack of an efficient transgenesis method. We have been developing transposon techniques by using the madaka fish Tol2 element. Taking advantage of its ability to generate genome-wide insertions, we developed the Gal4 gene trap and enhancer trap methods in zebrafish that enabled us to create various transgenic fish expressing Gal4 in specific cells. The Gal4-expressing cells can be visualized and manipulated in vivo by crossing the transgenic Gal4 lines with transgenic lines carrying various reporter and effector genes downstream of UAS (upstream activating sequence). Thus, the Gal4 gene trap and enhancer trap methods together with UAS lines now make detailed analyses of genes and cells in zebrafish feasible. Here, we describe the protocols to perform Gal4 gene trap and enhancer trap screens in zebrafish and their application to the studies of vertebrate neural circuits.
Genes / Markers
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Engineered Foreign Genes
Errata and Notes