Journal Article

The <i>Himar1 mariner</i> transposase cloned in a recombinant adenovirus vector is functional in mammalian cells

Linong Zhang, Uma Sankar, David J. Lampe, Hugh M. Robertson and Frank L. Graham

in Nucleic Acids Research

Volume 26, issue 16, pages 3687-3693
Published in print August 1998 | ISSN: 0305-1048
Published online August 1998 | e-ISSN: 1362-4962 | DOI: http://dx.doi.org/10.1093/nar/26.16.3687
The Himar1 mariner transposase cloned in a recombinant adenovirus vector is functional in mammalian cells

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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 two plasmids 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.

Journal Article.  4392 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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