Tol2-mediated Transgenesis, Gene Trapping, Enhancer Trapping, and the Gal4-UAS System

• Published in: Methods in Cell Biology Vol. 104; pp. 23 - 49
• Main Authors: Abe, Gembu, Suster, Maximilliano L., Kawakami, Koichi
• Format: Book Chapter Journal Article
• Language: English
• Published: United States Elsevier Science & Technology 2011
• Subjects: Animal breeding; Animals; Animals, Genetically Modified; Blotting, Southern - methods; Chromosomes, Artificial, Bacterial; Cloning, Molecular - methods; DNA Transposable Elements; Enhancer; Fishes (ichthyology); Gene trapping; Genes, Reporter; Germline; Green Fluorescent Proteins - biosynthesis; Green Fluorescent Proteins - genetics; Microinjection; Microinjections - methods; Mutagenesis, Insertional - methods; Recombination, Genetic; Transfection - methods; Transgenesis; Transposable; Zebrafish - genetics; Microinjection; Enhancer; Transposable; Transgenesis; Gene trapping; Germline
• ISBN: 9780123748140; 0123748143
• ISSN: 0091-679X
• DOI: 10.1016/B978-0-12-374814-0.00002-1

The Tol2 transposable element was originally found in the genome of the Japanese medaka fish ( Oryzias latipes). Tol2 contains a gene encoding an active transposase that can catalyze DNA transposition in vertebrate cells. In zebrafish, Tol2 generates genomic integrations in the germ cells very efficiently. By using the Tol2 transposition system, we have developed important genetic methods including transgenesis, gene trapping, enhancer trapping, and the Gal4-UAS system in zebrafish. In this chapter, we describe how these methods can be performed.