ZFIN ID: ZDB-PUB-090302-32
Temporally-controlled site-specific recombination in zebrafish
Hans, S., Kaslin, J., Freudenreich, D., and Brand, M.
Date: 2009
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
MeSH Terms:
  • Animals
  • Immunohistochemistry
  • In Situ Hybridization
  • Recombination, Genetic*
  • Zebrafish/genetics*
PubMed: 19247481 Full text @ PLoS One
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ABSTRACT
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.
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