The Frog Prince: a reconstructed transposon from Rana pipiens with high transpositional activity in vertebrate cells.
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 approximately 90% identical to a predicted transposase gene sequence from Xenopus laevis, but contained an unpredicted, approximately 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 approximately 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 approximately 70% higher activity in zebrafish cells than SB. Frog Prince can greatly extend our possibilities for genetic analyses in vertebrates.
Miskey
Csaba
C
Max Delbrück Center for Molecular Medicine, Robert Rössle Strasse 10, D-13092 Berlin, Germany.
Izsvák
Zsuzsanna
Z
Plasterk
Ronald H
RH
Ivics
Zoltán
Z
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D016384:Consensus Sequence / Q000235:genetics; D006224:Cricetinae; D004251:DNA Transposable Elements / Q000235:genetics*; D005399:Fishes / Q000235:genetics; D018628:Gene Dosage; D005818:Genetic Engineering; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010802:Phylogeny; D011894:Rana pipiens / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics*; D019895:Transposases / Q000235:genetics*; D014714:Vertebrates / Q000235:genetics*
D004251:DNA Transposable Elements; D019895:Transposases