| 新型RNAi载体抑制HPV16E6基因在人宫颈癌细胞Caski中的表达 |
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| 引用本文： | 杨峥嵘,何飞,李蓉,王海峰.新型RNAi载体抑制HPV16E6基因在人宫颈癌细胞Caski中的表达[J].生物磁学,2011(1):41-43. |
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| 作者姓名： | 杨峥嵘 何飞 李蓉 王海峰 |
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| 作者单位： | [1]深圳市疾病预防控制中心艾滋病实验室,广东深圳518055 [2]清华大学深圳研究生院生物医药研究中心,深圳518055 [3]深圳市人民医院口腔医学中心,广东深圳518020 [4]第三军医大学复合伤研究所,重庆400037 |
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| 基金项目： | 中国博士后基金项目（20060390073）; 深圳市科技计划项目 |
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| 摘 要： | 目的：利用自行构建的一种基于Semliki森林病毒的新型RNAi载体pSFV-RNAi Ready,验证将其用于短期高效沉默HPV16E6基因的效果。方法：以HPV16E6为靶标基因,设计并构建基于pSFV-RNAi Ready的重组质粒,分直接电击转染和病毒颗粒共培养两种方式转入人宫颈癌细胞株Caski,RT-PCR、Western blot检测HPV16E6表达水平。结果：重组质粒对HPV16E6沉默效果优于常规RNAi质料载体,接近化学合成小RNA,抑制率可高达90%以上,10天后效果仍然存在;结论：新型RNAi载体pSFV-RNAi Ready可较好地应用于特异高丰度靶基因的表达抑制,有望用于未来的科学研究或治疗应用。
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| 关 键 词： | RNA干扰 Semliki森林病毒 病毒载体 宫颈癌 |
A novel viral vector-mediated RNAi of HPV16 E6 expression in human cervical cancer cell Caski |
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| YANG Zheng-rong,HE Fei,LI Rong,WANG Hai-feng.A novel viral vector-mediated RNAi of HPV16 E6 expression in human cervical cancer cell Caski[J].Biomagnetism,2011(1):41-43. |
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| Authors: | YANG Zheng-rong HE Fei LI Rong WANG Hai-feng |
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| Institution: | 1 Department of AIDS Prevention,Shenzhen Center for Disease Control and Prevention,Shenzhen 518055,China; 2 Center for Biotech ＆ BioMedicine,Graduate School at Shenzhen,Tsinghua University,Shenzhen 518055,China; 3 Department of stomatology,people＇s hospital of Shenzhen,Shenzhen 518020,China; 4 Institute of Combined injury,Third Military Medical University,Chongqing 400037,China) |
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| Abstract: | Objective： To examine the silence effects of HPV 16E6 gene,which mediated by pSFV-RNAi Ready,a novel RNAi vector based on SFV replicator.Methods： Targeting HPV16E6 gene,Construct an re-combined plamid with pSFV-RNAi Ready,Then introduced into Caski cells by PLUS transfection and co-culture with viral particle.The expression of HPV16 E6 were investigated by RT-PCR and Western blot.Results： The novel pSFV-RNAi Ready vector was constructed successfully,Results of RT-PCR and Western blot indicated that pSFV-RNAi Ready mediated RNAi reduced HPV16E6 expression by more than 90 percent,which similar to chemi-cally synthesized short interfering RNAs,much stronger than traditional RNAi vector,and continue to work after more than 10 days.Conclusion： pSFV-RNAi Ready vector could silence the specific high-expressed target gene efficiently,It have potential for scientific research or clinlical therapy in the futrue. |
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| Keywords: | RNAi Semliki forest virus vector cervical cancer |
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