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

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Abstract
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.