| 杨峥嵘1,2 何飞3 李蓉4 王海峰2.新型RNAi 载体抑制HPV16E6 基因在人宫颈癌细胞Caski 中的表达[J].,2011,11(1):41-43 |
| 新型RNAi 载体抑制HPV16E6 基因在人宫颈癌细胞Caski 中的表达 |
| A novel viral vector-mediated RNAi of HPV16 E6 expression in humancervical cancer cell Caski |
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| DOI： |
| 中文关键词: RNA 干扰 Semliki 森林病毒 病毒载体 宫颈癌 |
| 英文关键词: RNAi Semliki forest virus vector cervical cancer |
| 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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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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 chemically
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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