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Nucleic Acids Research, 2003, Vol. 31, No. 23 6873-6881
© 2003 Oxford University Press

Article

The Frog Prince: a reconstructed transposon from Rana pipiens with high transpositional activity in vertebrate cells

Csaba Miskey1, Zsuzsanna Izsvák1,2, Ronald H. Plasterk3 and Zoltán Ivics*,1

1 Max Delbrück Center for Molecular Medicine, Robert Rössle Straße 10, D-13092 Berlin, Germany, 2 Institute of Biochemistry, Biological Research Center of the Hungarian Academy of Sciences, Szeged, Hungary and 3 Hubrecht Laboratory of Developmental Biology, Utrecht, The Netherlands

*To whom correspondence should be addressed. Tel: +49 30 9406 2546; Fax: +49 30 9406 2547; Email: zivics{at}mdc-berlin.de

Members of the Tc1/mariner superfamily of transposable elements isolated from vertebrates are transpositionally inactive due to the accumulation of mutations in their transposase genes. A novel open reading frame-trapping method was used to isolate uninterrupted transposase coding regions from the genome of the frog species Rana pipiens. The isolated clones were ~90% identical to a predicted transposase gene sequence from Xenopus laevis, but contained an unpredicted, ~180 bp region encoding the N-terminus of the putative transposase. None of these native genes was found to be active. Therefore, a consensus sequence of the transposase gene was derived. This engineered transposase and the transposon inverted repeats together constitute the components of a novel transposon system that we named Frog Prince (FP). FP has only ~50% sequence similarity to Sleeping Beauty (SB), and catalyzes efficient cut-and-paste transposition in fish, amphibian and mammalian cell lines. We demonstrate high-efficiency gene trapping in human cells using FP transposition. FP is the most efficient DNA-based transposon from vertebrates described to date, and shows ~70% higher activity in zebrafish cells than SB. Frog Prince can greatly extend our possibilities for genetic analyses in vertebrates.


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