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

Nucleic Acids Res. 1995 Oct 11;23(19):3816-21. doi: 10.1093/nar/23.19.3816.

Abstract

A recombinant adenovirus (Ad) expressing Cre recombinase derived from bacteriophage P1 was constructed. To assay the Cre activity in mammalian cells, another recombinant Ad bearing an on/off-switching reporter unit, where a LacZ-expression unit can be activated by the Cre-mediated excisional 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 establishing a powerful on/off switching strategy of gene expression in cultured 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Base Sequence
  • Blotting, Southern
  • Cell Line
  • DNA Nucleotidyltransferases / genetics*
  • Escherichia coli / genetics
  • Gene Expression Regulation*
  • Haplorhini
  • HeLa Cells
  • Humans
  • Integrases*
  • Kidney
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Recombinant Proteins / metabolism
  • T-Lymphocytes
  • Transcriptional Activation
  • Transfection
  • Viral Proteins*
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • Recombinant Proteins
  • Viral Proteins
  • Cre recombinase
  • DNA Nucleotidyltransferases
  • Integrases
  • beta-Galactosidase