中国现代医学杂志2017，Vol.27Issue(7)：30-35,6.DOI:10.3969/j.issn.1005-8982.2017.07.007
DNA载体介导的小干扰RNA Tet-on基因表达系统的调控效应检测
Detection of regulatory effects of a DNA vector-mediated Doxycycline-inducible siRNA system
摘要
Abstract
Objective To detect the regulatory effects of the established DNA vector-mediated Doxycycline-inducible siRNA system. Methods A DNA vector-mediated Doxycycline-inducible siRNA system, pDIRS4.1, was established. The regulatory effects of pDIRS4.1 were investigated by detecting the expression changes of enhanced green fluorescent protein (EGFP) and interleukin 6 (IL-6) after pDIRS4.1/EGFP siRNA, pEGFP-N3 plasmid, pDIRS4.1/IL-6 siRNA and pIRES IL-6 plasmid had been cotransfected into U-2OS Tet-On cells respectively which were added with serial concentrations of Doxycycline (Dox). Results The results showed that a high transfection efficiency (70%~80%) was obtained in the U-2OS Tet-On cells with pDIRS4.1 siRNA. pDIRS4.1 siRNA showed tight gene regulatory effects in the U-2OS Tet-On cells with the increase of Dox concentrations. Furthermore, the expressions of EGFP and IL-6 mediated by pDIRS4.1/EGFP siRNA and pDIRS4.1/IL-6 siRNA respectively had dose-dependent effect with the concentration of Dox. When the concentration of Dox was high, the expression level of IL-6 decreased 12 times. Conclusions A Dox-inducible siRNA expression system has been established through a non-viral mammalian expression vector. Tightly-regulated inducible siRNA expression systems of this type will likely have wide application in exploring gene functions in numerous model systems.关键词
小分子干扰RNA/四环素调控系统/DNA载体/基因调控Key words
small interference RNA/Tet-On system/DNA vector/gene regulation分类
医药卫生引用本文复制引用
李朵璐,周玉冰,韩超,安琪,李峰,段震峰,张晓坚,阚全程..DNA载体介导的小干扰RNA Tet-on基因表达系统的调控效应检测[J].中国现代医学杂志,2017,27(7):30-35,6.基金项目
河南省优秀创新型科技团队医学科技攻关计划项目(No:201303013) （No:201303013）
