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piggyBac ‐mediated germline transformation in the beetle Tribolium castaneum
Author(s) -
Lorenzen M. D.,
Berghammer A. J.,
Brown S. J.,
Denell R. E.,
Klingler M.,
Beeman R. W.
Publication year - 2003
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1046/j.1365-2583.2003.00427.x
Subject(s) - biology , transposable element , enhancer , germline , genetics , transformation (genetics) , mutagenesis , genome , transgene , insertional mutagenesis , enhancer trap , computational biology , mutant , gene , gene expression
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.