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| 腺相关病毒介导的RNA干扰抑制HeLa细胞中CTCF的表达 | |
| 引用本文： | 郭威威,朱旭东,刘党生,黄培堂. 腺相关病毒介导的RNA干扰抑制HeLa细胞中CTCF的表达[J]. 生物技术通讯, 2006, 17(4): 503-505 |
| 作者姓名： | 郭威威 朱旭东 刘党生 黄培堂 |
| 作者单位： | 1. 军事医学科学院,生物工程研究所,北京,100071;沈阳药科大学,辽宁,沈阳,110016 2. 军事医学科学院,生物工程研究所,北京,100071 3. 沈阳药科大学,辽宁,沈阳,110016 |
| 摘 要： | 目的:利用RNA干扰(RNAi)稳定抑制HeLa细胞中CTCF的表达。方法:构建能够抑制转录因子CTCF表达的短发夹RNA(shRNA)的腺相关病毒载体,重组病毒感染HeLa细胞后挑取单克隆,用Western印迹法和实时荧光定量PCR检测CTCF的表达水平。结果:获得3株CTCF被显著抑制的HeLa细胞株,Western印迹法和实时荧光定量PCR结果均显示CTCF的表达水平被抑制,其中最明显的被下调76%。结论:shRNA病毒表达载体构建成功,HeLa细胞中CTCF的表达可被长期稳定地抑制。 |
| 关 键 词： | 短发夹结构RNA CTCF 腺相关病毒载体 转录因子 |
| 文章编号： | 1009-0002(2006)04-0503-03 |
| 收稿时间： | 2006-02-03 |
| 修稿时间： | 2006-02-03 |
Inhibition Effect of RNAi on CTCF Expression in HeLa Cell Line by Adeno-Associated Virus Vector |
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| GUO Wei-wei,ZHU Xu-dong,LIU Dang-sheng,HUANG Pei-tang. Inhibition Effect of RNAi on CTCF Expression in HeLa Cell Line by Adeno-Associated Virus Vector[J]. Letters in Biotechnology, 2006, 17(4): 503-505 | |
| Authors: | GUO Wei-wei ZHU Xu-dong LIU Dang-sheng HUANG Pei-tang |
| Affiliation: | 1. Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071; 2. Shengyang Pharmaceutical University, Shenyang 110016; China |
| Abstract: | Objective: To suppress CTCF expression stably by RNAi in HeLa cell line.Methods: A shRNA expression vector was constructed through recombinant adeno-associated virus(rAAV) against CTCF gene and was introduced to HeLa cell.Then CTCF expressing level was detected by Western blotting and real-time fluorescent quantitative PCR in the selected HeLa cells.Result: It was demonstrated that the CTCF expression was knocked down specially 76% in 3 infected HeLa cell clones.Conclusion: The expression of CTCF is successfully decreased in HeLa cell line and the obtained shRNA expression vector through rAAV may be used for other functional analysis of CTCF. |
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