Adenovirus vector-mediated doxycycline-inducible RNA interference.
RNA interference (RNAi) is a powerful tool for the knockdown of gene expression. Here, we report on the development of an adenovirus (Ad) vector-mediated doxycycline (Dox)-inducible small interfering RNA (siRNA) expression system. We used this siRNA system to control the expression of p53 and c-Myc in human cancer cells. Coinfection of Ad vectors containing the siRNA expression system under the control of the Dox-inducible H1 promoter and Ad vectors expressing a tetracycline repressor inhibited the expression levels of p53 and c-Myc in a dose-dependent manner with both Dox and viral dose. Regulated silencing of p53 and c-Myc expression was obtained. Because an Ad vector-mediated inducible RNAi system can efficiently transduce a variety of cell types in vitro and in vivo, and the degree of loss of gene expression can be modulated according to the dose of Dox, this expression system should be a useful tool for both basic research on the analysis of gene function and therapeutic applications of RNAi.
Hosono
Tetsuji
T
Division of Cellular and Gene Therapy Products, National Institute of Health Sciences, Tokyo 158-8501, Japan.
Mizuguchi
Hiroyuki
H
Katayama
Kazufumi
K
Xu
Zhi-Li
ZL
Sakurai
Fuminori
F
Ishii-Watabe
Akiko
A
Kawabata
Kenji
K
Yamaguchi
Teruhide
T
Nakagawa
Shinsaku
S
Mayumi
Tadanori
T
Hayakawa
Takao
T
D000256:Adenoviridae / Q000235:genetics*; D015152:Blotting, Northern; D015153:Blotting, Western; D017931:DNA Primers; D004305:Dose-Response Relationship, Drug; D004318:Doxycycline / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D017931:DNA Primers; C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D004318:Doxycycline