Nucleic Acids Research, 1995, Vol. 23, No. 19 3816-3821
© 1995
MOLECULAR BIOLOGY |
Efficient gene activation in mammalian cells by using recombinant adenovirus expressing site-specific Cre recombinase
1Laboaory of Molecular Genetics University of Tokyo 4-6-1 Shirokanedi, Minato-ku, Tokyo 108, Japan 2Deparment of Virology, institute of Medical Science, University of Tokyo 4-6-1 Shirokanedai, Minato-ku, Tokyo 108, Japan 3Discovery Research Laboratories III, Sumitomo Pharmaceuticals Research Center 3-1-98 Kasugade-naka, Konohana-ku, Osaka 554, Japan
*To whom correspondence should be addressed
Received August 8, 1995. Accepted August 23, 1995.
A recombinant adenovirus (Ad) expressing Cre recombinase derived from bacteriophage P1 was constructed. To assay the Cre activity in mammalian colls, another recombinant Ad bearing an on/off-switching reporter unit, where a LacZ-expression unit can be activated by the Cre-mediated exclsionai deletion of an interposed stuffer DNA, was also constructed. Co-infection experiments together with the Cre-expressing and the reporter recombinant Ads showed that the Cre-mediated switching of gene expression was detected in nearly 100% of cultured CV1, HeLa and Jurkat cells. These results suggest that the recombinant Ad efficiently expressed functional Cre and offers a basis for estabilshing a powerful on/off switching strategy of gene expression incultured mammalian cells and presumably in transgenic animals. The method is also applicable to construction of recombinant Ad bearing a gene the expression of which is deleterious to propagation of recombinant Ad.
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