Résumé :
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Communication n° 181 Improvement of a non viral gene transfer strategy Amelioration of cis-acting elements of transposon gene transfer vector derived from MosI. F. Bonnin-Rouleux, S. Bigot, G. Jegot and Y. Bigot Laboratoire d'Etude des Parasites Génétiques EA3868 . UFR des Sciences et Techniques. Parc Grandmont, 37 200 Tours. France The mariner Mos I transposon element is especially well suited for use as a tool for transgene vectorisation. The cut and past transposable elements transpose by direct excision of the transgene from the vector and stable integration in the host genome.These elements function autonomously with minimal requirements for host cells functions and depend mainly on the fixation of the transposase to specific transposon sequences : inverted terminal repeat (ITR) and untranslated terminal repeat (UTR). The developement of a synthetic drive system based on Mos I have been hampered by low rate of integration. Optimisation of these sequences is one point critical for transposon application technology. We proposed to select new ITR and to study the impact of the UTR presence, in in vivo prokaryotic context : E. coli or in eukaryotic context : HeLa cells. These tests rely on the presence in the cells of the helper plasmid which encodes the transposase and the donor plasmid which encodes antibiotic resistance gene. The apparition of the new phenotype allow us to estimate the transposition efficiency and allow us to study the impact of the ameliorations. Up to now, there are a 17 and a 6 fold increase effect on the transposition frequency resulting from the use of new ITR as well as those observed by the presence of the UTR, in bacteria, respectively. The evaluation in HeLa cells are under investigation. This strategy was tested in the case of the project named SyntheGeneTransfer, that uses vectors made from MosI Mariner transposon to develop non-viral gene transfer.
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