Resetting the site: redirecting integration of an insertion sequence in a predictable way.
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.
Guynet
Catherine
C
Laboratoire de Microbiologie et Génétique Moléculaires, CNRS UMR5100, 118 Rte de Narbonne, F31062 Toulouse Cedex, France.
Achard
Adeline
A
Hoang
Bao Ton
BT
Barabas
Orsolya
O
Hickman
Alison Burgess
AB
Dyda
Frederick
F
Chandler
Michael
M
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*
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004277:DNA, Single-Stranded; D019895:Transposases