| RNA干扰抑制小鼠OX40基因表达的实验研究 |
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| 引用本文： | 夏仁品,卢实春,武聚山,李宁,郑静.RNA干扰抑制小鼠OX40基因表达的实验研究[J].免疫学杂志,2008,24(6):674-677. |
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| 作者姓名： | 夏仁品 卢实春 武聚山 李宁 郑静 |
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| 作者单位： | 首都医科大学附属北京佑安医院外科;上海吉凯基因化学技术有限公司; |
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| 基金项目： | 2007年教育部留学回国人员科研启动基金资助;;首都医科大学基础与临床合作基金(2006JL55)资助 |
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| 摘 要： | 目的研究靶向小鼠OX40基因的二聚体小干扰RNA(siRNA-OX40)对靶基因表达的沉默作用。方法建立并筛选稳定表达OX40基因的293T转染细胞株,用脂质体转染法将体外合成的siRNA-OX40转染上述细胞株,采用实时荧光定量PCR检测靶基因OX40 mRNA表达情况。结果用不同浓度的siRNA-OX40转染细胞48h,10nmol/L浓度对293T细胞靶基因表达的抑制作用最强(抑制率76.4%),1nmol/L浓度仍有一定的抑制作用(39.4%);25、10nmol/L siRNA转染293T细胞后,均在24h内起效(24.9%,20.5%),抑制作用至少能维持72h(74.4%,38.8%)。结论siRNA-OX40对293T细胞外源性靶基因表达有较强的特异性沉默作用,siRNA在24h内起效,48～72h达高峰,抑制作用至少能维持72h。本研究结果为下一步在动物或临床进行siRNA干扰试验提供了实验依据。
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| 关 键 词： | OX40 基因沉默 小干扰RNA |
Interference of OX40 gene expression induced by small interfering RNA |
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| XIA Ren-pin,LU Shi-chun,WU Ju-shan,LI Ning,ZHENG Jing.Interference of OX40 gene expression induced by small interfering RNA[J].Immunological Journal,2008,24(6):674-677. |
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| Authors: | XIA Ren-pin LU Shi-chun WU Ju-shan LI Ning ZHENG Jing |
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| Institution: | Department of Surgery;Beijing You-An Hospital;Capital Medical University;Beijing 100069;China |
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| Abstract: | Objective To investigate the specific interference of OX40 gene expression induced by RNAi technique in 293 T cell lines transfected with mice OX40 gene. Methods 293 T cells were transfected with recombined plasmid pEGFP-N1-GFP/ OX40,and then the positive cell clones were selected by fluorescence protein observation and RT-PCR. One specific dicer siRNA targeted to OX40 mRNA was designed and synthesized,sharing no homology with exons of known human gene. Quantitative real-time PCR was performed to measure th... |
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| Keywords: | OX40 |
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