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Genomic dsRNA of Grass carp reovirus protected from both the trigger and effector nucleases of RNA interference pathway during viral replication
Guo Shuai,Lu Liqun * #
College of Fisheries and Life Science, Shanghai Ocean University
*Correspondence author
#Submitted by
Subject:
Funding: Specialized Research Fund for the Doctoral Program of Higher Education of China（No.No.20093104120003）
Opened online: 2 April 2011
Accepted by: none
Citation: Guo Shuai,Lu Liqun.Genomic dsRNA of Grass carp reovirus protected from both the trigger and effector nucleases of RNA interference pathway during viral replication[OL]. [ 2 April 2011] http://en.paper.edu.cn/en_releasepaper/content/4418341
 
 
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
 
 
 

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