Nucleic Acids Research, 2001, Vol. 29, No. 7 e40
© 2001 Oxford University Press
Efficient gene activation in cultured mammalian cells mediated by FLP recombinase-expressing recombinant adenovirus
1Laboratory of Molecular Genetics, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan and 2Department of Biological Science and Technology, Science University of Tokyo, 2641 Yamazaki, Noda-shi, Chiba 278-8510, Japan
A recombinant adenovirus (rAd) expressing Cre recombinase derived from bacteriophage P1 has already been extensively used for the conditional gene activation and inactivation strategies in mammalian systems. In this study, we generated AxCAFLP, a rAd expressing FLP recombinase derived from Saccharomyces cerevisiae and carried out quantitative comparisons with Cre-expressing rAd in both in vitro and in cultured cells to provide another efficient gene regulation system in mammalian cells. In the in vitro experiments, the relative recombination efficiency of FLP expressed in 293 cells infected with FLP-expressing rAd was approximately one-thirtieth that of Cre even at 30°C, the optimum temperature for FLP activity, and was approximately one-ninetieth at 37°C. Co-infection experiments in HeLa cells using a target rAd conditionally expressing LacZ under the control of FLP showed that an FLP-expressing rAd, infected at a multiplicity of infection (MOI) of 5, was able to activate the transgene in almost 100% of HeLa cells whereas the Cre-expressing rAd was sufficient at an MOI of 0.2. Since an MOI of 5 is ordinarily used in rAd experiments, these results showed that the FLP-expressing rAd is useful for gene activation strategies and is probably applicable to a sequential gene regulation system in combination with Cre-expressing rAd in mammalian cells.
* To whom correspondence should be addressed. Tel: +81 3 5449 5627; Fax: +81 3 5449 5432; Email: isaito{at}ims.u-tokyo.ac.jp Present address:Kazuhiko Odaka, Marketing Planning Group, Marketing Department, Research Products Division, Daiichi Pure Chemicals Co. Ltd, 2-8-3 Higashinihonbashi, Chuo-ku, Tokyo 103-0004, Japan
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