piggyBac-mediated germline transformation in the beetle Tribolium castaneum

Insect Mol Biol. 2003 Oct;12(5):433-40. doi: 10.1046/j.1365-2583.2003.00427.x.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Southern
  • Chromosome Mapping
  • DNA Transposable Elements / genetics*
  • Gene Expression Profiling
  • Microinjections
  • Molecular Sequence Data
  • Plasmids
  • Transformation, Genetic*
  • Tribolium / genetics*

Substances

  • DNA Transposable Elements