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Background  The  Sleeping Beauty  ( SB ) transposon system has been used for germline transgenesis of the diploid frog,  Xenopus tropicalis . Injecting one-cell embryos with plasmid DNA harboring an  SB  transposon substrate together with mRNA encoding the  SB  transposase enzyme resulted in non-canonical integration of small-order concatemers of the transposon. Here, we demonstrate that  SB  transposons stably integrated into the frog genome are effective substrates for remobilization.    Results  Transgenic frogs that express the  SB 10 transposase were bred with  SB  transposon-harboring animals to yield double-transgenic 'hopper' frogs. Remobilization events were observed in the progeny of the hopper frogs and were verified by Southern blot analysis and cloning of the novel integrations sites. Unlike the co-injection method used to generate founder lines, transgenic remobilization resulted in canonical transposition of the  SB  transposons. The remobilized  SB  transposons frequently integrated near the site of the donor locus; approximately 80% re-integrated with 3 Mb of the donor locus, a phenomenon known as 'local hopping'.    Conclusions  In this study, we demonstrate that  SB  transposons integrated into the  X. tropicalis  genome are effective substrates for excision and re-integration, and that the remobilized transposons are transmitted through the germline. This is an important step in the development of large-scale transposon-mediated gene- and enhancer-trap strategies in this highly tractable developmental model system.

Remobilization of Sleeping Beauty transposons in the germline of Xenopus tropicalis