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Mol Cell. 2009 Jun 12;34(5):612-9. doi: 10.1016/j.molcel.2009.05.017.

Resetting the site: redirecting integration of an insertion sequence in a predictable way.

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  • 1Laboratoire de Microbiologie et Génétique Moléculaires, CNRS UMR5100, 118 Rte de Narbonne, F31062 Toulouse Cedex, France.

Abstract

Target site choice is a complex and poorly understood aspect of DNA transposition despite its importance in rational transposon-mediated gene delivery. Though most transposons choose target sites essentially randomly or with some slight sequence or structural preferences, insertion sequence IS608 from Helicobacter pylori, which transposes using single-stranded DNA, always inserts just 3' of a TTAC tetranucleotide. Our results from studies on the IS608 transposition mechanism demonstrated that the transposase recognizes its target site by co-opting an internal segment of transposon DNA and utilizes it for specific recognition of the target sites through base-pairing. This suggested a way to redirect IS608 transposition to novel target sites. As we demonstrate here, we can now direct insertions in a predictable way into a variety of different chosen target sequences, both in vitro and in vivo.

PMID:
19524540
[PubMed - indexed for MEDLINE]
PMCID:
PMC3654794
Free PMC Article

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