piggyBac can bypass DNA synthesis during cut and paste transposition

Rupak Mitra, Jennifer Fain-Thornton, Nancy L. Craig

Research output: Contribution to journalArticlepeer-review

145 Scopus citations

Abstract

DNA synthesis is considered a defining feature in the movement of transposable elements. In determining the mechanism of piggyBac transposition, an insect transposon that is being increasingly used for genome manipulation in a variety of systems including mammalian cells, we have found that DNA synthesis can be avoided during piggyBac transposition, both at the donor site following transposon excision and at the insertion site following transposon integration. We demonstrate that piggyBac transposon excision occurs through the formation of transient hairpins on the transposon ends and that piggyBac target joining occurs by the direct attack of the 3′OH transposon ends on to the target DNA. This is the same strategy for target joining used by the members of DDE superfamily of transposases and retroviral integrases. Analysis of mutant piggyBac transposases in vitro and in vivo using a piggyBac transposition system we have established in Saccharomyces cerevisiae suggests that piggyBac transposase is a member of the DDE superfamily of recombinases, an unanticipated result because of the lack of sequence similarity between piggyBac and DDE family of recombinases.

Original languageEnglish (US)
Pages (from-to)1097-1109
Number of pages13
JournalEMBO Journal
Volume27
Issue number7
DOIs
StatePublished - Apr 9 2008

Keywords

  • Precise excision
  • Transposase
  • Transposition
  • piggyBac

ASJC Scopus subject areas

  • General Neuroscience
  • Molecular Biology
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

Fingerprint

Dive into the research topics of 'piggyBac can bypass DNA synthesis during cut and paste transposition'. Together they form a unique fingerprint.

Cite this