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 from the horn fly, is independent of host-specific factors that would limit transfer between different species, making it an ideal candidate for gene transfer technology development. To determine the activity of Himar1 transposase in mammalian cells, we introduced the Himar1 transposase gene into an adenovirus (Ad) vector under control 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 transposition reactions, Himar1 transposase expressed by the Ad vector mediated precise cut-and-paste transposition and resulted in a characteristic duplication of TA at the integration site of the target plasmid. Further studies showed that this transposase was capable of catalyzing transposition between twoplasmids co-transfected into 293T7pol cells, which express T7 RNA polymerase. Combining the integration capability of mariner transposons with the transduction efficiency of Ad vectors is expected to provide a powerful tool for introducing transgenes into the host chromosome.
Department of Biology and Department of Pathology, McMaster University, Hamilton, Ontario L8S 4K1, Canada.
L
L
Zhang
U
U
Sankar
D J
DJ
Lampe
H M
HM
Robertson
F L
FL
Graham
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004274:DNA, Recombinant / Q000235:genetics; D004268:DNA-Binding Proteins; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D008957:Models, Genetic; D015345:Oligonucleotide Probes / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D019895:Transposases / Q000235:genetics* / Q000378:metabolism*
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; C111652:mariner transposases; D019895:Transposases