PUBLICATION
Advanced Zebrafish Transgenesis with Tol2 and Application for Cre/lox Recombination Experiments
- Authors
- Mosimann, C., and Zon, L.I.
- ID
- ZDB-PUB-110921-12
- Date
- 2011
- Source
- Methods in cell biology 104: 173-194 (Chapter)
- Registered Authors
- Mosimann, Christian, Zon, Leonard I.
- Keywords
- Tol2, trangenesis, transgenes, Cre, CreERT2, IoxP, lineage tracing, tamoxifen, 4-OHT
- MeSH Terms
-
- Animals
- Animals, Genetically Modified*
- Cloning, Molecular/methods
- DNA Transposable Elements*
- Female
- Genetic Markers
- Genetic Testing/methods
- Genetic Vectors/isolation & purification
- Integrases/genetics
- Integrases/metabolism
- Male
- Microinjections/methods
- Mutagenesis, Insertional/methods*
- Recombinant Proteins/genetics
- Recombinant Proteins/metabolism
- Recombination, Genetic*
- Viral Proteins/genetics
- Viral Proteins/metabolism
- Zebrafish/genetics*
- PubMed
- 21924163 Full text @ Meth. Cell. Biol.
Citation
Mosimann, C., and Zon, L.I. (2011) Advanced Zebrafish Transgenesis with Tol2 and Application for Cre/lox Recombination Experiments. Methods in cell biology. 104:173-194.
Abstract
Spatio-temporal transgene regulation by transgenic DNA recombinases is a central tool for genetic research in multicellular organisms, with excellent applications for misexpression and lineage tracing experiments. Cre recombinase-controlled lox site recombination is a cornerstone of contemporary mouse genetics, and Cre/lox techniques therefore attract increasing interest in the zebrafish field. Tol2-mediated zebrafish transgenesis now provides a stable platform for lox cassette transgenes, while the ease of drug treatments in zebrafish makes the model an ideal candidate for Tamoxifen/4-hydroxytamoxifen-inducible CreER(T2) experiments. In this chapter, we will first introduce the basics of Cre/lox methodology, CreER(T2) regulation by Tamoxifen/4-hydroxytamoxifen, as well as the benefits of Tol2 transgenesis for Cre/lox experiments. We will then in detail outline practical experimental steps for Tol2 transgenesis toward the creation of single-insertion transgenes. Lastly, we will introduce protocols for 4-hydroxytamoxifen-mediated CreER(T2) induction to perform spatio-temporal lox transgene regulation experiments in zebrafish embryos.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping