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, 1995•academic.oup.comA 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 …
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