piggyBac-mediated germline transformation in the beetle Tribolium castaneum.
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.
Lorenzen
M D
MD
Division of Biology, Kansas State University, Manhattan, KS, USA.
Berghammer
A J
AJ
Brown
S J
SJ
Denell
R E
RE
Klingler
M
M
Beeman
R W
RW
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*
D004251:DNA Transposable Elements