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PLoS One. 2009;4(2):e4640. doi: 10.1371/journal.pone.0004640. Epub 2009 Feb 27.

Temporally-controlled site-specific recombination in zebrafish.

Author information

1
Biotechnology Center and Center for Regenerative Therapies Dresden, Dresden University of Technology, Dresden, Germany.

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.

PMID:
19247481
PMCID:
PMC2645673
DOI:
10.1371/journal.pone.0004640
[Indexed for MEDLINE]
Free PMC Article

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