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Nucleic Acids Research, 2003, Vol. 31, No. 11 2795-2802
© 2003 Oxford University Press

Faithful expression of a tagged Fugu WT1 protein from a genomic transgene in zebrafish: efficient splicing of pufferfish genes in zebrafish but not mice

Colin G. Miles, Lesley Rankin, Shirley I. Smith, Martina Niksic, Greg Elgar1 and Nicholas D. Hastie

Medical Research Council Human Genetics Unit, Western General Hospital, Creve Road, Edinburgh EH4 2XU, UK and 1 United Kingdom Human Genome Mapping Project Resource Centre, Cambridge CB10 1SB, UK

*To whom correspondence should be addressed. Tel: +44 131 332 2471; Fax: +44 131 467 8424; Email: nick.hastie{at}hgu.mrc.ac.uk

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 ß-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.


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