Post-integration stabilization of a transposon vector by terminal sequence deletion in Drosophila melanogaster

Nat Biotechnol. 2004 Sep;22(9):1150-4. doi: 10.1038/nbt1002. Epub 2004 Aug 8.

Abstract

Germline transformation systems for nearly 20 insect species have been derived from transposable elements, allowing the development of transgenic insects for basic and applied studies. These systems use a defective nonautonomous vector that results in stable vector integrations after the disappearance of transiently provided transposase helper plasmid, which is essential to maintain true breeding lines and consistent transgene expression that would otherwise be lost after vector remobilization. The risk of remobilization by an unintended transposase source has so far not been a concern for laboratory studies, but the prospective use of millions of transgenic insects in biocontrol programs will likely increase the risk, therefore making this a critical issue for the ecological safety of field release programs. Here we describe an efficient method that deletes a terminal repeat sequence of a transposon vector after genomic integration. This procedure prevents transposase-mediated remobilization of the other terminal sequence and associated genes, ensuring their genomic stability.

Publication types

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

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics*
  • DNA Transposable Elements / genetics*
  • Drosophila melanogaster / genetics*
  • Gene Deletion*
  • Genetic Vectors / genetics*
  • Genomic Instability / genetics*
  • Mutagenesis, Site-Directed / genetics
  • Terminal Repeat Sequences / genetics*

Substances

  • DNA Transposable Elements