短发夹状RNA在人细胞诱导RNA干扰(英文)

Short Hairpin RNAs Induced RNA Interference in Human Cells

背景与目的:RNA干扰是一种通过细胞内导入双链RNA而导致特异基因表达抑制的进化保守的转录后基因沉默现象,目前RNA干扰已成为研究基因功能的重要方法。本研究旨在利用DNA载体在细胞内产生短发夹状RNA(shorthairpinRNAs,shRNA),通过这些shRNA来诱导RNA干扰(RNAinterference,RNAi),为基因功能分析提供新的实验手段。方法:利用双荧光素酶报告系统来检测DNA载体在细胞内产生的shRNA诱导RNAi的效果。比较该载体产生的shRNA在不同条件下的RNA干扰效果。结果:DNA载体产生的shRNA在人细胞能诱导RNAi,序列特异地抑制基因表达。抑制效果与所选基因靶位点高度相关。结论:shRNA在人细胞能诱导RNA干扰,序列特异地抑制基因表达,这一方法可用于基因功能分析。

BACKGROUND &OBJECTIVE:RNA interference(RNAi )is an evolutionarily conserved posttran scriptional g ene silencing ,in whic h the introduction of double-stranded RN A into a cell leads to specific suppre ssion of g ene expression.RNAi has become an i mportant tool for g ene function stud ies.The aim of this study was to induce RNAi in mammalian cells by short hairpin RNAs (shRNAs )g enerated from a DNA vector,therefo re,to provide a new approach for g ene function analysis.METHODS :Dual Luciferase System was used to assess the intracellular eff ect of the shRNAs g enerated from a DNA vector.The inhibitory effect of thi s DNA vector-driven shRNAs at differ ent condition was measured.RESULTS :shRNAs g enerated from the DNA vector induced RNAi in human cells ,and suppressed g ene expression in a sequence-specific manner.The inhi bitory effect was hig hly related to t he targ et sites.CONCLUSION:shRNAs induced RNAi in human cells an d suppressed g ene expression in a sequence-speci fic manner.This approach may be used for g ene function study.