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

• 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/

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.