title: piggyBac-mediated germline transformation in the beetle Tribolium castaneum.
abstract: The lepidopteran transposable element piggyBac can mediate germline insertions in at least four insect orders. It therefore shows promise as a broad-spectrum transformation vector, but applications such as enhancer trapping and transposon-tag mutagenesis are still lacking. We created, cloned, sequenced and genetically mapped a set of piggyBac insertions in the red flour beetle, Tribolium castaneum. Transpositions were precise, and specifically targeted the canonical TTAA recognition sequence. We detected several novel reporter-expression domains, indicating that piggyBac could be used to identify enhancer regions. We also demonstrated that a primary insertion of a non-autonomous element can be efficiently remobilized to non-homologous chromosomes by injection of an immobile helper element into embryos harbouring the primary insertion. These developments suggest potential for more sophisticated methods of piggyBac-mediated genome manipulation.
authors.affiliation: Division of Biology, Kansas State University, Manhattan, KS, USA.
authors.forename: M D
authors.initials: MD
authors.lastname: Lorenzen
authors.forename: A J
authors.initials: AJ
authors.lastname: Berghammer
authors.forename: S J
authors.initials: SJ
authors.lastname: Brown
authors.forename: R E
authors.initials: RE
authors.lastname: Denell
authors.forename: M
authors.initials: M
authors.lastname: Klingler
authors.forename: R W
authors.initials: RW
authors.lastname: Beeman
mesh_terms: D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002874:Chromosome Mapping; D004251:DNA Transposable Elements / Q000235:genetics*; D020869:Gene Expression Profiling; D008845:Microinjections; D008969:Molecular Sequence Data; D010957:Plasmids; D014170:Transformation, Genetic; D014231:Tribolium / Q000235:genetics*
chemical_list: D004251:DNA Transposable Elements