THE HIMAR1 MARINER TRANSPOSASE CLONED IN A RECOMBINANT ADENOVIRUS VECTOR IS FUNCTIONAL IN MAMMALIAN-CELLS

Mariner transposons belong to the mariner/Tc1 superfamily of class II, DNA-mediated elements. One of these transposons, Himar1, isolated fro m the horn fly, is independent of host-specific factors that would lim it transfer between different species, making it an ideal candidate fo r gene transfer technology development. To determine the activity of H imar1 transposase in mammalian cells, we introduced the Himar1 transpo sase gene into an adenovirus (Ad) vector under central of the phage T7 RNA polymerase promoter. Mammalian cells infected with the Ad vector carrying the Himar1 gene efficiently expressed the Himar1 transposase in the presence of T7 polymerase, In in vitro inter-plasmid transposit ion reactions, Himar1 transposase expressed by the Ad vector mediated precise cut-and-paste transposition and resulted in a characteristic d uplication of TA at the integration site of the target plasmid, Furthe r studies showed that this transposase was capable of catalyzing trans position between two plasmids co-transfected into 293T7pol cells, whic h express T7 RNA polymerase, Combining the integration capability of m ariner transposons with the transduction efficiency of Ad vectors is e xpected to provide a powerful tool for introducing transgenes into the host chromosome.