The bacteriophage T7 binary system activates transient transgene expression in zebrafish (Danio rerio) embryos.
The bacteriophage T7 binary expression system is widely used in vitro for high level selective expression of cloned genes but its application to in vivo models has not yet been investigated. In the present work, we show that coinjection into fertilized zebrafish eggs of pE1T7R, an expression plasmid bearing the T7 RNA polymerase gene driven by the cytomegalovirus (CMV) promoter, together with reporter vectors containing the Escherichia coli lacZ gene driven by the T7 promoter, resulted in the efficient expression of the reporter gene in 24-h mosaic transgenic embryos. Conversely, embryos receiving an unrelated CMV-expression plasmid, instead of pE1T7R, lacked significant reporter gene activity, indicating the strict requirement of T7 polymerase to activate the T7 promoter in these embryos. The present study demonstrates the possibility of applying efficiently the bacteriophage T7 binary system in vivo to a vertebrate model.
Verri
T
T
Department of Biology, University of Padua, Italy.
Argenton
F
F
Tomanin
R
R
Scarpa
M
M
Storelli
C
C
Costa
R
R
Colombo
L
L
Bortolussi
M
M
D000818:Animals; D030801:Animals, Genetically Modified; D017123:Bacteriophage T7 / Q000235:genetics*; D003587:Cytomegalovirus / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000096:biosynthesis / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000502:physiology*; D004926:Escherichia coli; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014764:Viral Proteins; D015027:Zebrafish; D001616:beta-Galactosidase / Q000096:biosynthesis*
D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D001616:beta-Galactosidase