title: A novel conditional gene silencing method using a tumor-specific and heat-inducible siRNA system.
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.
authors.affiliation: Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Feng
authors.affiliation: Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liao
authors.affiliation: Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Feng
authors.affiliation: Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, China. tangliling@cqu.edu.cn.
authors.forename: Liling
authors.initials: L
authors.lastname: Tang
mesh_terms: D001483:Base Sequence; D045744:Cell Line, Tumor; D020868:Gene Silencing; D018390:Gene Targeting; D005822:Genetic Vectors; D018869:Heat-Shock Response; D056945:Hep G2 Cells; D006358:Hot Temperature; D006801:Humans; D061986:MCF-7 Cells; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D009369:Neoplasms / Q000235:genetics* / Q000628:therapy; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics*; D051845:gamma-Synuclein / Q000235:genetics / Q000378:metabolism
chemical_list: D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; C104249:SNCG protein, human; D051845:gamma-Synuclein
keywords: Gene silence; HSE; HTERT promoter; Heat-shock; SiRNA