首页|期刊导航|中华医学杂志（英文版）|Potent and specific inhibition of SARS-CoV antigen expression by RNA interference
中华医学杂志（英文版）2005，Vol.118Issue(9)：714-719,6.
Potent and specific inhibition of SARS-CoV antigen expression by RNA interference
Potent and specific inhibition of SARS-CoV antigen expression by RNA interference
摘要
Abstract
Background Severe acute respiratory syndrome (SARS) is an infectious disease caused by SARS-CoV. There are no effective antiviral drugs for SARS although the epidemic of SARS was controlled. The aim of this study was to develop an RNAi (RNA interference) approach that specifically targeted the N gene sequence of severe acute respiratory syndrome associated coronavirus (SARS-CoV) by synthesizing short hairpin RNA (shRNA) in vivo, and to assess the inhibitory effect of this shRNA on SARS-CoV N antigen expression. Methods The eukaryotic expression plasmid pEGFP-C1-N, containing SARS-CoV N gene, was co-transfected into 293 cells with either the RNAi plasmid pshRNA-N or unrelated control plasmid pshRNA-HBV-C4. At 24, 48 and 72 hours post transfection, the green fluorescence was observed through a fluorescence microscope. The RNA levels of SARS-CoV N were determined by reverse transcription polymerase chain reaction (RT-PCR). The expression of Green Fluorescent Protein (GFP) and protein N were detected using Western blot.Results The vector, pshRNA-N expressing shRNA which targeted the N gene of SARS-CoV, was successfully constructed. The introduction of RNAi plasmid efficiently and specifically inhibited the synthesis of protein N. RT-PCR showed that RNAs of N gene were clearly reduced when the pEGFP-C1-N was cotransfected with pshRNA-N, whereas the control vector did not exhibit inhibitory effect on N gene transcription.Conclusions Our results demonstrate that RNAi mediated silencing of SARS-CoV gene could effectively inhibit expression of SARS-CoV antigen, hence RNAi based strategy should be further explored as a more efficacious antiviral therapy of SARS-CoV infection.关键词
RNA interference/severe acute respiratory syndrome/virus/gene therapyKey words
RNA interference/severe acute respiratory syndrome/virus/gene therapy分类
医药卫生引用本文复制引用
TAO Peng,ZHANG Jun,TANG Ni,ZHANG Bing-qiang,HE Tong-chuan,HUANG Ai-long..Potent and specific inhibition of SARS-CoV antigen expression by RNA interference[J].中华医学杂志（英文版）,2005,118(9):714-719,6.基金项目
This work was supported by the grants from National Science Fund for Distinguished Young Scholars (No.30228026) and National 863 Program in the Tenth Five-Year Plan Period (No.2001AA217121). （No.30228026）
