文献类型：期刊文献

中文题名：光诱导RNAi系统的构建及优化

英文题名：Optimization and Construction of Light-Inducible RNAi System

作者：王建华[1];王雪[1];杜增民[1];赵玉政[1];杨弋[1]

机构：[1]华东理工大学药学院生物反应器工程国家重点实验室,上海200237

年份：2015

卷号：41

期号：1

起止页码：28

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；CSCD:【CSCD2015_2016】；

基金：国家自然科学基金(31225008;31170815;31071260);上海市科委"创新行动计划"基础研究重点项目(NO12JC1402900);上海市曙光人才计划(11SG31);高等学校博士学科点专项科研基金(20100074110010);高等学校基本科研业务费

语种：中文

中文关键词：RNAi;光诱导基因表达系统;shRNAmir;萤火虫荧光素酶

外文关键词：RNAi; light-inducible gene expression system; shRNAmir; firefly luciferase

摘要：利用光诱导基因表达系统,构建光诱导RNAi系统,并对系统转染比例、转录因子的表达量和shRNAmir表达载体的核心启动子进行优化。通过构建不同启动子的光敏转录因子GAVPO表达载体和不同核心启动子的shRNAmir表达载体,分析它们对报告基因的干扰效果的影响。结果表明,GAVPO的表达量对报告基因沉默效果和shRNAmir的背景表达水平至关重要。低表达量的GAVPO能够实现对报告基因的微调控,报告基因的表达量下降到约60%;而高表达量的GAVPO能够高效抑制报告基因表达,报告基因的表达量下降到34%。
A light-inducible RNAi system was built by using the light-inducible gene expression system.The transfection mass ratio,the expression level of transcription factors and the core promoter of shRNAmir expression vector were optimized.We constructed transcription factor GAVPO expression vectors containing different promoter segments and the shRNAmir expression vectors with different core promoters,and assayed their effects on inhibition of gene expression.The results show that the GAVPO expression level plays an important role in gene silencing and background expression level of shRNAmir.Low expression level of GAVPO can tightly control expression of reporter gene,and reporter gene expression level decreased to about 60%;overexpression of GAVPO can efficiently inhibit the reporter gene expression,and reporter gene expression level decreased to 34%.