Temporally-controlled site-specific recombination in zebrafish.
Conventional 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.
Hans
Stefan
S
Biotechnology Center and Center for Regenerative Therapies Dresden, Dresden University of Technology, Dresden, Germany.
Kaslin
Jan
J
Freudenreich
Dorian
D
Brand
Michael
M
D000818:Animals; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D011995:Recombination, Genetic; D015027:Zebrafish / Q000235:genetics*