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. 2012 Apr 11;27(2):109–119. doi: 10.1007/s12250-012-3230-4

Suppression of RNA interference pathway in vitro by Grass carp reovirus

Shuai Guo 1, Dan Xu 1, Hong-xu Xu 2, Tu Wang 1, Jia-le Li 1, Li-qun Lu 1,✉
PMCID: PMC8218150  PMID: 22492002

Abstract

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.

Key words: Grass carp reovirus (GCRV), RNA interference (RNAi), Dicer, Northern blot

Footnotes

Foundation items: The Shanghai committee of Science and Technology (Grant No.10PJ1404800) and the National Natural Science Foundation of China (Grant No. 31072244).

These authors contributed equally to this work.

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