Efficient gene activation in mammalian cells by using recombinant adenovirus expressing site-specific Cre recombinase

Y Kanegae, G Lee, Y Sato, M Tanaka… - Nucleic Acids …, 1995 - academic.oup.com
Y Kanegae, G Lee, Y Sato, M Tanaka, M Nakal, T Sakaki, S Sugano, I Saito
Nucleic Acids Research, 1995academic.oup.com
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 …
Abstract
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.
Oxford University Press