PUBLICATION

Tol2-mediated transgenesis, gene trapping, enhancer trapping, and Gal4-UAS system

Authors
Kawakami, K., Asakawa, K., Muto, A., Wada, H.
ID
ZDB-PUB-160725-34
Date
2016
Source
Methods in cell biology   135: 19-37 (Chapter)
Registered Authors
Asakawa, Kazuhide, Kawakami, Koichi, Muto, Akira, Wada, Hironori
Keywords
Enhancer trapping, Gene trapping, The Gal4-UAS system, The Gal4FF trans-activator, The Tol2 transposable element, Transgenesis
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • DNA Transposable Elements/genetics*
  • Enhancer Elements, Genetic*
  • Gene Transfer Techniques*
  • Germ Cells/growth & development
  • Oryzias/genetics
  • Plasmids/genetics
  • Zebrafish/genetics*
  • Zebrafish/growth & development
PubMed
27443919 Full text @ Meth. Cell. Biol.
Abstract
The Tol2 element is an active transposon that was found from the genome of the Japanese medaka fish. Since the Tol2 transposition system is active in all vertebrate cells tested so far, it has been applied to germ line transgenesis in various model animals including fish, frog, chicken, and mouse, and to gene transfer in culture cells. In zebrafish, the Tol2 system consists of the transposase mRNA and a Tol2 transposon-donor plasmid, and is introduced into fertilized eggs by microinjection. Thus genomic integrations of the Tol2 construct are generated in the germ lineage and transmitted to the offspring very efficiently. By using the Tol2 transposition system, we have developed important genetic methods, such as transgenesis, gene trapping, enhancer trapping, and the Gal4-UAS system in zebrafish and applied to many aspects of biological studies. In this chapter, we describe how these methods are performed.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping