CAJ | 학술논문

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( short hairpin RNAs,shRNA) ,通过这些 shRNA来诱导 RNA干扰( RNA interference,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.
배경여목적: RNA간우시일충통과세포내도입쌍련 RNA이도치특이기인표체억제적진화보수적전록후기인침묵현상,목전 RNA간우이성위연구기인공능적중요방법.본연구지재이용 DNA재체재세포내산생단발협상 RNA( short hairpin RNAs,shRNA) ,통과저사 shRNA래유도 RNA간우( RNA interference,RNAi),위기인공능분석제공신적실험수단.방법:이용쌍형광소매보고계통래검측 DNA재체재세포내산생적 shRNA유도 RNAi적효과.비교해재체산생적 shRNA재불동조건하적 RNA간우효과.결과: DNA재체산생적 shRNA재인세포능유도 RNAi,서렬특이지억제기인표체.억제효과여소선기인파위점고도상관.결론: shRNA재인세포능유도 RNA간우,서렬특이지억제기인표체,저일방법가용우기인공능분석.
BACKGROUND ＆ OBJECTIVE:RNA interference(RNAi) is an evolutionarily conserved posttranscriptional gene silencing, in which the introduction of double- stranded RNA into a cell leads to specific suppression of gene expression. RNAi has become an important tool for gene function studies. The aim of this study was to induce RNAi in mammalian cells by short hairpin RNAs(shRNAs) generated from a DNA vector, therefore, to provide a new approach for gene function analysis. METHODS: Dual Luciferase System was used to assess the intracellular effect of the shRNAs generated from a DNA vector. The inhibitory effect of this DNA vector- driven shRNAs at different condition was measured. RESULTS: shRNAs generated from the DNA vector induced RNAi in human cells, and suppressed gene expression in a sequence- specific manner. The inhibitory effect was highly related to the target sites. CONCLUSION: shRNAs induced RNAi in human cells and suppressed gene expression in a sequence- specific manner. This approach may be used for gene function study.