Ubiquitous transgene expression and Cre-based recombination driven by the ubiquitin promoter in zebrafish.
Molecular genetics approaches in zebrafish research are hampered by the lack of a ubiquitous transgene driver element that is active at all developmental stages. Here, we report the isolation and characterization of the zebrafish ubiquitin (ubi) promoter, which drives constitutive transgene expression during all developmental stages and analyzed adult organs. Notably, ubi expresses in all blood cell lineages, and we demonstrate the application of ubi-driven fluorophore transgenics in hematopoietic transplantation experiments to assess true multilineage potential of engrafted cells. We further generated transgenic zebrafish that express ubiquitous 4-hydroxytamoxifen-controlled Cre recombinase activity from a ubi:cre(ERt2) transgene, as well as ubi:loxP-EGFP-loxP-mCherry (ubi:Switch) transgenics and show their use as a constitutive fluorescent lineage tracing reagent. The ubi promoter and the transgenic lines presented here thus provide a broad resource and important advancement for transgenic applications in zebrafish.
Howard Hughes Medical Institute, Boston, MA 02115, USA.
Christian
C
Mosimann
Charles K
CK
Kaufman
Pulin
P
Li
Emily K
EK
Pugach
Owen J
OJ
Tamplin
Leonard I
LI
Zon
D000818:Animals; D030801:Animals, Genetically Modified; D004789:Enzyme Activation / Q000187:drug effects; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology; D019076:Transgenes / Q000235:genetics*; D025801:Ubiquitin / Q000235:genetics*; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism*
D025801:Ubiquitin; D013629:Tamoxifen; C016601:afimoxifene; C045073:Cre recombinase; D019426:Integrases