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串联序列amiRNA稳定转染后对cccDNA的影响

Influence of the tandem sequence expression plasmids of amiRNA against HBV cccDNA
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摘要 目的:观察优化设计的串联序列amiRNA(artificial microRNA)质粒表达载体长期干扰乙型肝炎病毒(HBV)后,对HBV复制源泉cccDNA的影响。方法:将前期构建的针对HBV RNA干扰效率高的单一序列及串联序列amiRNA质粒表达载体,瞬时转染入HepG2.2.15细胞,筛选出稳定转染细胞株,观察其对cccDNA的影响。结果:稳定转染单一序列amiRNA-HBV-4组相对于阴性对照质粒,对HBV cccDNA的抑制率为93.78%;串联序列amiRNA-HBV3-4组为99.65%,二者比较,差异有高度统计学意义(P<0.01)。结论:构建的针对HBV的新型amiRNA质粒表达载体可较明显地抑制HBV复制的根源cccDNA,串联序列组效果更佳,为进一步进行体内实验奠定了基础。 Objective: To observe the effect of cccDNA after the long-term inhibition of tandem sequence amiRNA plasraids expression vector against HBV. Methods: The earlier constructed amiRNA plasmids of singular and tandem sequences against HBV were transfected into HepG2.2.15 cells transiently. Then the cells which stably expressed miRNA were screened. Last cccDNA were measured to observe the long-term inhibition efficiency. Results: HBV cccDNA remarkably decreased in amiRNA-HBV-4 group by 93.78% compared with the negative control plasmid group; amiRNA- HBV3-4 group by 99.65%, there was a significant difference between them (P〈0.01). Conclusion: The tandem sequence plasmid expression vector amiRNA can remarkably inhibit replication of HBV cccDNA, and the tandem sequence group is better. It pave the way for this plasmid expressing carrier to further study in vivo.
出处 《中国医药导报》 CAS 2011年第31期20-23,F0003,共5页 China Medical Herald
基金 辽宁省大连市科技局科技计划项目(编号:2010E15SF168)
关键词 乙型肝炎病毒(HBV) amiRNA 串联序列质粒表达载体 稳定转染 CCCDNA HBV amiRNA Tandem sequence plasmids expression vector Stable transfection cccDNA
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参考文献15

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