Nucleic Acids Research, 2001, Vol. 29, No. 4 e23
© 2001 Oxford University Press
A novel tetracycline-dependent transactivator with E2F4 transcriptional activation domain
Saitama Cancer Center Research Institute, 818 Komuro Ina Kita-adachigun Saitama 362-0806, Japan, 1Department of Tumor Genetics and Biology, Kumamoto University School of Medicine, 2-2-1 Honjo, Kumamoto 860-0811, Japan and 2Division of Molecular Genetics and Center of Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands
A tetracycline-controlled gene expression system provides a powerful tool to dissect the functions of gene products. However, it often appears difficult to establish cell lines or transgenic animals stably expressing tetracycline-dependent transactivators, possibly as a result of toxicity of the transactivator domains used. In order to overcome this problem, we developed a novel tetracycline-dependent transactivator that works efficiently in mammalian cells. This transactivator is a fusion of the tet reverse repressor mutant and the transcriptional activating domain of human E2F4, which is ubiquitously expressed in vivo. We demonstrate here that this tetracycline-regulated gene expression system provides a two log transcriptional activation in mammalian cells as assessed by northern blot and luciferase analyses. Combining this system with green fluorescent protein reporter systems or microarray gene expression profiling will facilitate the study of gene function.
* To whom correspondence should be addressed. Tel: +81 48 722 1111; Fax: +81 48 722 1739; Email: akagi{at}cancer-c.pref.saitama.jp Present address: Masayuki Kanai, Department of Biochemistry and Molecular Oncology, Institute of Basic Medical Sciences, University of Tsukuba, 1-1-1 Tennoudai Tsukuba Ibaraki 305-8575, Japan