For example, the ET7 line harbors a transposon insertion near the mkp3 locus and expresses GFP in the midbrain-hindbrain boundary, forebrain and the ventricle, matching a subset of the known FGF8-dependent mkp3 expression domain. We have identified both previously characterized as well as novel expression patterns from this screen. Many of the insertions are into introns of zebrafish genes predicted in the current genome assembly. We have molecularly characterized these lines and show that in each line, a specific GFP expression pattern is due to a single transposition event. We established 9 lines of zebrafish with distinct tissue- or organ-specific GFP expression patterns from 90 founders that produced GFP-expressing progeny. We modulated a Sleeping Beauty transposon-based transgenesis cassette to establish an enhancer trapping technique for use in a vertebrate model system, zebrafish Danio rerio. Pioneering experiments in Drosophila have demonstrated the value of enhancer "trapping" using an invertebrate to address this functional genomics problem. Among functional elements of a metazoan gene, enhancers are particularly difficult to find and annotate.
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