A novel conditional gene silencing method using a tumor-specific and heat-inducible siRNA system. (1st June 2016)

• Record Type: Journal Article
• Authors: Feng, Jing; Wang, Xiaoyu; Liao, Yi; Feng, Jianguo; Tang, Liling
• Is Part Of: Journal of industrial microbiology & biotechnology. Volume 43:Number 6(2016)
• Page Start: 761
• Page End: 770
• Publication Date: 2016-06-01
• Subjects: HTERT promoter -- HSE -- SiRNA -- Heat-shock -- Gene silence; Industrial microbiology -- Periodicals; 660.62
• DOI: 10.1007/s10295-016-1759-1
• Languages: English
• ISSNs: 1367-5435

Abstract

Abstract: RNAi technology is an invaluable tool for investigating gene function. However, the non-temporal and non-spatial control is the primary limitation, which leads to siRNA leakiness and off-target effects. In this study, we inserted three kinds of HSE into tumor specific promoter hTERT, which aims to construct a temperature-inducible and tumor-specific RNAi plasmid vector. In our system, the expression of mature siRNA is tightly controlled by the heat shock-inducible and tumor-specific promoters. From the expression level of RNA and protein, we determined the efficiency of the inducible siRNA system by targeting SNCG gene in HepG2 and MCF-7 cells. Results showed that the controllable siRNA system could be induced to initiate siRNA expression by heat-induce. The silencing effect of SNCG is on a relative low level (10 %) at 37 °C, while it is significantly increased to 50 or 60 % after heat inducing at 43 °C. This new conditional siRNA system provides a novel approach to drive the siRNA expression by heat-inducible and tumor-specific promoter.