） 摘要： 目的：RNAi起始于宿主细胞Dicer蛋白特异性地结合并且切割双链RNA
本文以草鱼呼肠孤病毒GCRV为模型探讨GCRV复制过程中其双链RNA基因组逃逸宿主RNAi信号通路的作用机制
吕利群
呼肠孤病毒的复制抑制RNAi信号通路中核酸酶对其双链RNA基因组的降解[EB/OL]
北京：中国科技论文在线
并进一步转染GCRV感染的CIK细胞研究GCRV的复制对RNAi信号通路的影响
结论：GCRV的复制不但能抑制引发RNAi的Dicer核酸酶对其基因组的切割
结果：GCRV的复制感染可以诱导Dicer蛋白的基因的转录水平升高
） Abstract： Objective: The present study aimed to investigate the effect of GCRV replication on Dicer-initiated RNAi pathway of host cell
Methods: The pEGFP-N1 was transfected into Grass carp kidney cells (CIK) with either the synthetic EGFP-speific siRNA or in intro transcribed EGFP-specific dsRNA to signify the functional gene silencing pathway in CIK cell line
Then, CIK cells were infected with GCRV and the steady-state transcription levels of Dicer were monitored during viral replication cycle using real-time quantitative PCR (qRT-PCR)
The effect of GCRV replication on RNA interference pathway was further studied by investigating the silencing of egfp gene by egfp-siRNA in GCRV-infected CIK cells
Results: both egfp-siRNA and egfp-dsRNA could silence the egfp reporter gene in uninfected CIK cells
In GCRV-infected cells, replication of GCRV correlated with the increased transcription level of Dicer gene while resulted in the loss function of egfp-siRNA on silencing reporter egfp gene
Conclusion: Replication of GCRV augmented the transcription of Dicer gene but still antagonized the classical RNAi pathway of host cells
Suppression of the siRNA-induced RNAi by GCRV replication suggested that GCRV could inhibit the effector protein of RNAi pathway besides the insensitivity of its dsRNA genome to Dicer nuclease, the RNAi- trigger nuclease
Keywords： Grass carp reovirus (GCRV); RNA interference (RNAi); Dicer 下载PDF阅读器 PDF全文下载： 初稿 （ 158 ） 作者简介： Guo Shuai, (1986-), Female, Graduate student, Virology 通信联系人： Lu Liqun,Professor, Virology 【收录情况】 中国科技论文在线： 郭帅
GCRV不是通过抑制Dicer的转录表达逃逸Dicer的切割
方法：将pEGFP-N1质粒和egfp基因特异性的siRNA或dsRNA共转染草鱼CIK细胞验证RNAi途径在CIK细胞中的存在
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