title: Resetting the site: redirecting integration of an insertion sequence in a predictable way.
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.
authors.affiliation: Laboratoire de Microbiologie et Génétique Moléculaires, CNRS UMR5100, 118 Rte de Narbonne, F31062 Toulouse Cedex, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Guynet
authors.forename: Adeline
authors.initials: A
authors.lastname: Achard
authors.forename: Bao Ton
authors.initials: BT
authors.lastname: Hoang
authors.forename: Orsolya
authors.initials: O
authors.lastname: Barabas
authors.forename: Alison Burgess
authors.initials: AB
authors.lastname: Hickman
authors.forename: Frederick
authors.initials: F
authors.lastname: Dyda
authors.forename: Michael
authors.initials: M
authors.lastname: Chandler
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D020029:Base Pairing; D001483:Base Sequence; D004251:DNA Transposable Elements / Q000502:physiology*; D004277:DNA, Single-Stranded / Q000737:chemistry*; D016480:Helicobacter pylori / Q000235:genetics*; D008957:Models, Genetic; D017354:Point Mutation; D019895:Transposases / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004277:DNA, Single-Stranded; D019895:Transposases