dc.contributor.author | van der Wal, Yorick Andreas | |
dc.contributor.author | Nordli, Henriette R. | |
dc.contributor.author | Akandwanaho, Allan | |
dc.contributor.author | Greiner-Tollersrud, Linn | |
dc.contributor.author | Kool, Jaap | |
dc.contributor.author | Jørgensen, Jorunn B | |
dc.date.accessioned | 2023-10-16T07:14:37Z | |
dc.date.available | 2023-10-16T07:14:37Z | |
dc.date.issued | 2023-07-17 | |
dc.description.abstract | Background: Interferon (IFN) responses are critical in the resolution of viral
infections and are actively targeted by many viruses. They also play a role in
inducing protective responses after vaccination and have been successfully
tested as vaccine adjuvants. IFN responses are well conserved and function
very similar in teleosts and mammals. Like in mammals, IFN responses in piscine
cells are initiated by intracellular detection of the viral infection by different
pattern recognition receptors. Upon the recognition of viral components, IFN
responses are rapidly induced to combat the infection. However, many viruses
may still replicate and be able to inhibit or circumvent the IFN response by
different means.<p>
<p>Methods: By employing CRISPR Cas9 technology, we have disrupted proteins
that are central for IFN signaling in the salmonid cell line CHSE-214. We
successfully generated KO clones for the mitochondrial antiviral signaling
protein MAVS, the transcription factors IRF3 and IRF7-1, as well as a double KO
for IRF7-1/3 using an optimized protocol for delivery of CRISPR-Cas
ribonucleoproteins through nucleofection.
<p>Results: We found that MAVS and IRF3 KOs inhibited IFN and IFN-stimulated
gene induction after intracellular poly I:C stimulation as determined through
gene expression and promoter activation assays. In contrast, the IRF7-1 KO had
no clear effect. This shows that MAVS and IRF3 are essential for initiation of
intracellular RNA-induced IFN responses in CHSE-214 cells. To elucidate viral
interference with IFN induction pathways, the KOs were infected with Salmon
alphavirus 3 (SAV3) and infectious pancreatic necrosis virus (IPNV). SAV3 infection
in control and IRF7-1 KO cells yielded similar titers and no cytopathic effect, while
IRF3 and MAVS KOs presented with severe cytopathic effect and increased titers
6 days after SAV 3 infection. In contrast, IPNV yields were reduced in IRF3 and MAVS KOs, suggesting a dependency on interactions between viral proteins and
pattern recognition receptor signaling components during viral replication.
<p>Conclusion: Aside from more insight in this signaling in salmonids, our results
indicate a possible method to increase viral titers in salmonid cells. | en_US |
dc.identifier.citation | van der Wal, Nordli, Akandwanaho, Greiner-Tollersrud, Kool, Jørgensen. CRISPR-Cas– induced IRF3 and MAVS knockouts in a salmonid cell line disrupt PRR signaling and affect viral replication. Frontiers in Immunology. 2023;14 | en_US |
dc.identifier.cristinID | FRIDAID 2183929 | |
dc.identifier.doi | 10.3389/fimmu.2023.1214912 | |
dc.identifier.issn | 1664-3224 | |
dc.identifier.uri | https://hdl.handle.net/10037/31567 | |
dc.language.iso | eng | en_US |
dc.publisher | Frontiers Media | en_US |
dc.relation.journal | Frontiers in Immunology | |
dc.rights.accessRights | openAccess | en_US |
dc.rights.holder | Copyright 2023 The Author(s) | en_US |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0 | en_US |
dc.rights | Attribution 4.0 International (CC BY 4.0) | en_US |
dc.title | CRISPR-Cas– induced IRF3 and MAVS knockouts in a salmonid cell line disrupt PRR signaling and affect viral replication | en_US |
dc.type.version | publishedVersion | en_US |
dc.type | Journal article | en_US |
dc.type | Tidsskriftartikkel | en_US |
dc.type | Peer reviewed | en_US |