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

Type Article
Date 2017-10-18
Language English
Author(s) Pauletto Marianna1, Segarra Amelie2, Montagnani Caroline3, Quillien Virgile4, Faury Nicole2, Le Grand Jacqueline4, Miner Philippe4, Petton Bruno4, Labreuche Yannick5, Fleury ElodieORCID4, Fabioux Caroline6, Bargelloni Luca1, Renault Tristan7, Huvet ArnaudORCID4
Affiliation(s) 1 : Univ Padua, Dept Comparat Biomed & Food Sci, Viale Univ 16, I-35020 Legnaro, PD, Italy.
2 : IFREMER, Lab Genet & Pathol Mollusques Marins, F-17390 La Tremblade, France.
3 : Univ Montpellier, IFREMER, Univ Perpignan Via Domitia, IHPE UMR 5244,CNRS, F-34095 Montpellier, France.
4 : UBO, UMR CNRS 6539, IFREMER, IRD,LEMAR, F-29280 Plouzane, France.
5 : UPMC Paris 06, Sorbonne Univ, CNRS, UMR 8227,Stn Biol Roscoff, CS 90074, F-29688 Roscoff, France.
6 : Univ Bretagne Occidentale, UMR CNRS 6539, IFREMER, Inst Univ Europeen Mer,IRD,LEMAR, F-29280 Plouzane, France.
7 : IFREMER, Dept Ressources Biol & Environm, Rue lIle dYeu, F-44000 Nantes, France.
Source Journal Of Experimental Biology (0022-0949) (Company Of Biologists Ltd), 2017-10-18 , Vol. 220 , N. 20 , P. 3671-3685
DOI 10.1242/jeb.156299
WOS© Times Cited 3
Keyword(s) Anti-viral response, RNA interference, Inhibitor of NF-kappa B, Marine bivalve, Ostreid herpesvirus 1
Abstract

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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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 (2017). Long dsRNAs promote an anti-viral response in Pacific oyster hampering ostreid herpesvirus 1 replication. Journal Of Experimental Biology, 220(20), 3671-3685. Publisher's official version : https://doi.org/10.1242/jeb.156299 , Open Access version : https://archimer.ifremer.fr/doc/00409/52011/