Studies on vertebrate developmental genes by transposon-mediated gene trapping

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(1)We injected a transposon-donor plasmid containing a To12 construct and mRNA encoding the transposase into zebrafish fertilized eggs and demonstrated that the transposon construct can be integrated in the genome of the germ cells very efficiently. This method is now widely used as a standard method to create transgenic fish.(2)We constructed the To12 construct containing a splice acceptor and the GFP reporter gene and created random integration of this construct in the zebrafish genome. When the splice acceptor trapped a transcript of an endogenous gene, the GFP gene is expressed in a pattern similar to that of the endogenous gene. This was the first successful demonstration of gene trapping in zebrafish.(3)We have been making efforts to distribute this useful methodology by publication of manuscripts, talks in international meetings, writing review articles, a training course and collaboration.(4)In the course of the transposon studies in zebrafish, we also found that the To12 transposon system is also active in mouse and Xenopus. To12 will be a useful gene transfer vector in these model vertebrates.