title: Faithful expression of a tagged Fugu WT1 protein from a genomic transgene in zebrafish: efficient splicing of pufferfish genes in zebrafish but not mice.
abstract: The teleost fish are widely used as model organisms in vertebrate biology. The compact genome of the pufferfish, Fugu rubripes, has proven a valuable tool in comparative genome analyses, aiding the annotation of mammalian genomes and the identification of conserved regulatory elements, whilst the zebrafish is particularly suited to genetic and developmental studies. We demonstrate that a pufferfish WT1 transgene can be expressed and spliced appropriately in transgenic zebrafish, contrasting with the situation in transgenic mice. By creating both transgenic mice and transgenic zebrafish with the same construct, we show that Fugu RNA is processed correctly in zebrafish but not in mice. Furthermore, we show for the first time that a Fugu genomic construct can produce protein in transgenic zebrafish: a full-length Fugu WT1 transgene with a C-terminal beta-galactosidase fusion is spliced and translated correctly in zebrafish, mimicking the expression of the endogenous WT1 gene. These data demonstrate that the zebrafish:Fugu system is a powerful and convenient tool for dissecting both vertebrate gene regulation and gene function in vivo.
authors.affiliation: Medical Research Council Human Genetics Unit, Western General Hospital, Creve Road, Edinburgh EH4 2XU, UK.
authors.forename: Colin G
authors.initials: CG
authors.lastname: Miles
authors.forename: Lesley
authors.initials: L
authors.lastname: Rankin
authors.forename: Shirley I
authors.initials: SI
authors.lastname: Smith
authors.forename: Martina
authors.initials: M
authors.lastname: Niksic
authors.forename: Greg
authors.initials: G
authors.lastname: Elgar
authors.forename: Nicholas D
authors.initials: ND
authors.lastname: Hastie
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D030801:Animals, Genetically Modified; D015870:Gene Expression; D016678:Genome; D007438:Introns; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D013045:Species Specificity; D023741:Takifugu / Q000235:genetics*; D019076:Transgenes; D025721:WT1 Proteins / Q000096:biosynthesis / Q000235:genetics*; D015027:Zebrafish / Q000235:genetics*
chemical_list: D025721:WT1 Proteins