|ZFIN ID: ZDB-PUB-090302-32|
Temporally-controlled site-specific recombination in zebrafish
Hans, S., Kaslin, J., Freudenreich, D., and Brand, M.
|Source:||PLoS One 4(2): e4640 (Journal)|
|Registered Authors:||Brand, Michael, Freudenreich, Dorian, Hans, Stefan, Kaslin, Jan|
|Keywords:||Embryos, Zebrafish, Alleles, Recombinant proteins, In situ hybridization, Diencephalon, Gene mapping, Plasmid construction|
|PubMed:||19247481 Full text @ PLoS One|
Hans, S., Kaslin, J., Freudenreich, D., and Brand, M. (2009) Temporally-controlled site-specific recombination in zebrafish. PLoS One. 4(2):e4640.
ABSTRACTConventional use of the site-specific recombinase Cre is a powerful technology in mouse, but almost absent in other vertebrate model organisms. In zebrafish, Cre-mediated recombination efficiency was previously very low. Here we show that using transposon-mediated transgenesis, Cre is in fact highly efficient in this organism. Furthermore, temporal control of recombination can be achieved by using the ligand-inducible CreER(T2). Site-specific recombination only occurs upon administration of the drug tamoxifen (TAM) or its active metabolite, 4-hydroxy-tamoxifen (4-OHT). Cre-mediated recombination is detectable already 4 or 2 hours after administration of TAM or 4-OHT, demonstrating fast recombination kinetics. In addition, low doses of TAM allow mosaic labeling of single cells. Combined, our results show that conditional Cre/lox will be a valuable tool for both, embryonic and adult zebrafish studies. Furthermore, single copy insertion transgenesis of Cre/lox constructs suggest a strategy suitable also for other organisms.