Suppression of RNA interference pathway in vitro by Grass carp reovirus
Suppression of RNA interference pathway in vitro by Grass carp reovirus
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草鱼呼肠孤病毒对RNA干扰途径的体外抑制
DOI：
10.1007/s12250-012-3230-4
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发表时间：
2012-04-01
影响因子：
5.5
通讯作者：
Lu, Li-qun
中科院分区：
文献类型：
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作者：
Guo, Shuai;Xu, Dan;Lu, Li-qun
呼肠孤病毒基因组dsRNA从dsRNA触发和Dicer启动的RNAi途径中存活的方式仍有待确定。研究草鱼呼肠孤病毒（GCRV）复制对草鱼肾细胞（CIK）RNAi途径的影响。通过RNAi实验证明dsRNA触发的RNAi途径在CIK细胞中未受损。北方印迹分析显示，转染纯化的GCRV基因组dsRNA的CIK细胞中产生了GCRV特异性siRNA，而在GCRV感染的CIK细胞中未检测到GCRV特异性siRNA。感染和转染实验进一步表明GCRV的复制与Dicer基因转录水平的增加以及体外合成的egfp-siRNA在沉默EGFP报告基因中的功能抑制相关。这些数据表明，虽然只有GCRV的基因组dsRNA对细胞RNAi途径敏感，但未鉴定的RNAi抑制蛋白可能有助于病毒基因组的存活和有效的病毒复制。
The means of survival of genomic dsRNA of reoviruses from dsRNA-triggered and Dicer-initiated RNAi pathway remains to be defined. The present study aimed to investigate the effect of Grass carp reovirus (GCRV) replication on the RNAi pathway of grass carp kidney cells (CIK). The dsRNA-triggered RNAi pathway was demonstrated unimpaired in CIK cells through RNAi assay. GCRV-specific siRNA was generated in CIK cells transfected with purified GCRV genomic dsRNA in Northern blot analysis; while in GCRV-infected CIK cells, no GCRV-specific siRNA could be detected. Infection and transfection experiments further indicated that replication of GCRV correlated with the increased transcription level of the Dicer gene and functional inhibition of in vitro synthesized egfp-siRNA in silencing the EGFP reporter gene. These data demonstrated that although only the genomic dsRNA of GCRV was sensitive to the cellular RNAi pathway, unidentified RNAi suppressor protein(s) might contribute to the survival of the viral genome and efficient viral replication.