| 唐敏，熊晶.慢病毒载体对HIF-2α基因沉默效率的研究.[J].中南医学科学杂志.,2012,40(3):240-244. |
| 慢病毒载体对HIF-2α基因沉默效率的研究 |
| The Study on Silencing Efficiency of HIF-2α Gene in 293T Cells by SiRNA Slow Virus Expression Vector |
| 投稿时间：2012-03-09 |
| DOI： |
| 中文关键词: HIF-2α基因 慢病毒载体 RNA干扰 |
| 英文关键词:HIF-2α gene slow virus vector RNA interference |
| 基金项目: |
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| 中文摘要: |
| 目的构建有效抑制HIF-2α基因表达的siRNA序列，并找到最佳病毒载体浓度。方法设计针对HIF-2α mRNA作用的4条siRNA的序列，分别插入慢病毒载体中，获得较高滴度的慢病毒后，运用RT-PCR和Werstern blot方法检测HIF-2α mRNA及蛋白质水平，筛选出对HIF-2α基因沉默作用较强的干扰序列及病毒载体剂量。结果4条siRNA序列均能干扰HIF-2α基因表达，其中KD2序列对HIF-2α基因的干扰作用最强（P<0.05）。两种不同剂量的慢病毒载体比较，高剂量慢病毒载体的干扰效果更明显（P<0.05）。结论KD2序列能有效沉默HIF-2α基因；高剂量的慢病毒载体对HIF-2α基因的沉默效果更强。 |
| 英文摘要: |
| ObjectiveIn this study,by screening the four siRNA interference sequence,which consider HIF-2α mRNA as target sequences,we find the most effective way to inhibit expression vector of HIF-2α.And we find the best density of viral vector.MethodWe used RNAi design software to design role for HIF-2α mRNA in the four siRNA sequences,and inserted into the lentiviral vector respectively.We detected the four siRNA interference effect by using RT-PCR and Western blot.Then,we screened for interference sequence which had strong effect on HIF-2α gene silencing.ResultsIn the four siRNA sequences,KD2 sequence had the strongest effect on the expression of HIF-2α interference(P<0.05),and successfully constructed siRNA-KD2 lentivirus vector and obtained a higher titer of the virus.ConclusionIn the four siRNA sequences,KD2 sequence has the strongest effect on the expression of HIF-2α interference(P<0.05).KD2 can effectively inhibit the expression of HIF-2α gene.It can successfully construct siRNA-KD2 lentivirus vector and obtain a higher titer of the virus.The lentiviral vector PshRAN-copGFP-lentivector can effectively load for siRNA specific sequence for HIF-2α gene. |
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