Long dsRNAs promote an anti-viral response in Pacific oyster hampering ostreid herpesvirus 1 replication

Double-stranded RNA (dsRNA)-mediated genetic interference (RNAi) is a widely used reverse genetic tool for determining the loss-of-function phenotype of a gene. Here, the possible induction of an immune response by long dsRNA was tested in a marine bivalve (Crassostrea gigas), as well as the specifi...

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Bibliographic Details
Published in:Journal of Experimental Biology
Main Authors: Pauletto, Marianna, Segarra, Amelie, Montagnani, Caroline, Quillien, Virgile, Faury, Nicole, Le Grand, Jacqueline, Miner, Philippe, Petton, Bruno, Labreuche, Yannick, Fleury, Elodie, Fabioux, Caroline, Bargelloni, Luca, Renault, Tristan, Huvet, Arnaud
Format: Article in Journal/Newspaper
Language:English
Published: Company Of Biologists Ltd 2017
Subjects:
Anti-viral response
RNA interference
Inhibitor of NF-kappa B
Marine bivalve
Ostreid herpesvirus 1
Pacific
Crassostrea gigas
Pacific oyster
Online Access:https://archimer.ifremer.fr/doc/00409/52011/52698.pdf
https://doi.org/10.1242/jeb.156299
https://archimer.ifremer.fr/doc/00409/52011/
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Summary:Double-stranded RNA (dsRNA)-mediated genetic interference (RNAi) is a widely used reverse genetic tool for determining the loss-of-function phenotype of a gene. Here, the possible induction of an immune response by long dsRNA was tested in a marine bivalve (Crassostrea gigas), as well as the specific role of the subunit 2 of the nuclear factor kappa B inhibitor (I kappa B2). This gene is a candidate of particular interest for functional investigations in the context of oyster mass mortality events, as Cg-I kappa B2 mRNA levels exhibited significant variation depending on the amount of ostreid herpesvirus 1 (OsHV-1) DNA detected. In the present study, dsRNAs targeting Cg-I kappa B2 and green fluorescent protein genes were injected in vivo into oysters before being challenged by OsHV-1. Survival appeared close to 100% in both dsRNA-injected conditions associated with a low detection of viral DNA and a low expression of a panel of 39 OsHV-1 genes as compared with infected control. Long dsRNA molecules, both Cg-I kappa B2- and GFP-dsRNA, may have induced an anti-viral state controlling the OsHV-1 replication and precluding the understanding of the specific role of Cg-I kappa B2. Immune-related genes including Cg-I kappa B1, Cg-Rel1, Cg-IFI44, Cg-PKR and Cg-IAP appeared activated in the dsRNA-injected condition, potentially hampering viral replication and thus conferring a better resistance to OsHV-1 infection. We revealed that long dsRNA-mediated genetic interference triggered an anti-viral state in the oyster, emphasizing the need for new reverse genetics tools for assessing immune gene function and avoiding off-target effects in bivalves.

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