Antisense downregutation of SARS-CoV gene expression in Vero E6 cells
文献类型：期刊论文
| 作者 | Shi, Y; Luo, HF; Jia, J; Xiong, J; Yang, DH; Huang, B; Jin, YX |
| 刊名 | JOURNAL OF GENE MEDICINE
 |
| 出版日期 | 2005 |
| 卷号 | 7期号:1页码:97-107 |
| 关键词 | SARS antisense technology Vero E6 cells RT-PCR EGFP fusion protein |
| 通讯作者 | Jin, YX (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, Yueyang Rd 320, Shanghai 200031, Peoples R China.,yxjin@sibs.ac.cn |
| 英文摘要 | Background Severe acute respiratory syndrome (SARS) is caused by a novel coronavirus (SARS-CoV). It is an enveloped, single-stranded plus-sense RNA virus with a genome of similar to30 kb. The structural proteins E, M and N of SARS-CoV play important roles during host cell entry and vital morphogenesis and release. Therefore, we have studied whether expression of these structural proteins can be down-regulated using an antisense technique. Methods Vero E6 cells were transfected with plasmid constructs containing exons of the SARS-CoV structural protein E, M or N, genes or their exons in frame with the reporter protein EGFP. The transfected cell cultures were treated with antisense phosphorothioated oligonucleotides (antisense PS-ODN, 20mer) or a control oligonucleotide by addition to the culture medium. Results Among a total of 26 antisense PS-ODNs targeting E. N-and N 0 a. genes, we obtained six antisense PS-ODNs which could sequence-specifically reduce target genes expression by over 90% at the concentration of 50 mu,M in the cell culture medium tested by RT-PCR. The antisense effect was proved by down-regulating the expression of the fusion proteins containing the structural proteins E. M or N in frame with the reporter protein EGFP. In Vero E6 cells, the antisense effect was dependent on the concentrations of the antisense PS-ODNs in a range of 0-10 muM or 0-30 muM. Conclusions The antisense PS-ODNs are effective in downregulation of SARS. The findings indicate that antisense knockdown of SARS could be a useful strategy for treatment of SARS, and could also be suitable for studies of the pathological function of SARS genes in a cellular model system. Copyright (C) 2004 John Wiley Sons, Ltd. |
| 学科主题 | Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine |
| 类目[WOS] | Biotechnology & Applied Microbiology ; Genetics & Heredity ; Medicine, Research & Experimental |
| 关键词[WOS] | ACUTE RESPIRATORY SYNDROME ; HUMAN-IMMUNODEFICIENCY-VIRUS ; PHOSPHOROTHIOATE ANALOGS ; GENOME SEQUENCE ; CORONAVIRUS ; OLIGODEOXYNUCLEOTIDE ; REPLICATION ; OLIGONUCLEOTIDES ; INHIBITION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000226620500010 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1849]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Shi, Y,Luo, HF,Jia, J,et al. Antisense downregutation of SARS-CoV gene expression in Vero E6 cells[J]. JOURNAL OF GENE MEDICINE,2005,7(1):97-107. |
| APA | Shi, Y.,Luo, HF.,Jia, J.,Xiong, J.,Yang, DH.,...&Jin, YX.(2005).Antisense downregutation of SARS-CoV gene expression in Vero E6 cells.JOURNAL OF GENE MEDICINE,7(1),97-107. |
| MLA | Shi, Y,et al."Antisense downregutation of SARS-CoV gene expression in Vero E6 cells".JOURNAL OF GENE MEDICINE 7.1(2005):97-107. |
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