Cre-mediated site-specific recombination in zebrafish embryos.
Cre-mediated site-specific recombination has become an invaluable tool for manipulation of the murine genome. The ability to conditionally activate gene expression or to generate chromosomal alterations with this same tool would greatly enhance zebrafish genetics. This study demonstrates that the HSP70 promoter can be used to inducibly control expression of an enhanced green fluorescent protein (EGFP) -Cre fusion protein. The EGFP-Cre fusion protein is capable of promoting recombination between lox sites in injected plasmids or in stably inherited transgenes as early as 2 hr post-heat shock induction. Finally, the levels of Cre expression achieved in a transgenic fish line carrying the HSP70-EGFP-cre transgene are compatible with viability and both male and female transgenic fish are fertile subsequent to induction of EGFP-Cre expression. Hence, our data suggests that Cre-mediated recombination is a viable means of manipulating gene expression in zebrafish.
Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas, USA. thummel.1@nd.edu
Ryan
R
Thummel
Christopher T
CT
Burket
Jeffrey L
JL
Brewer
Michael P
MP
Sarras
Li
L
Li
Martin
M
Perry
Jeffrey P
JP
McDermott
Brian
B
Sauer
David R
DR
Hyde
Alan R
AR
Godwin
D000818:Animals; D030801:Animals, Genetically Modified; D004625:Embryo, Nonmammalian / Q000502:physiology; D017930:Genes, Reporter; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D019426:Integrases; D009030:Mosaicism; D016297:Mutagenesis, Site-Directed; D011993:Recombinant Fusion Proteins / Q000235:genetics; D011995:Recombination, Genetic; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D018840:HSP70 Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; C045073:Cre recombinase; D019426:Integrases; D001616:beta-Galactosidase