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However, flaviviruses sequester -vRNA into membrane-bound replication compartments away from cytosolic sensors. We found that single-stranded -vRNA accumulates outside of replication compartments in \u201cfirst responder\u201d cells, rendering it accessible to RLRs. Exposure of this -vRNA occurs at late timepoints of infection, is linked to viral assembly, and depends on the expression of viral structural proteins. These findings reveal that while most infected cells replicate high levels of vRNA, release of -vRNA from replication compartments during assembly occurs at low frequency and is critical for initiation of innate immunity during flavivirus infection.<\/jats:p>","DOI":"10.1101\/2024.06.07.597966","type":"posted-content","created":{"date-parts":[[2024,6,8]],"date-time":"2024-06-08T17:10:10Z","timestamp":1717866610000},"source":"Crossref","is-referenced-by-count":0,"title":["Exposure of negative-sense viral RNA in the cytoplasm initiates innate immunity to West Nile virus"],"prefix":"10.64898","author":[{"given":"Emmanuelle","family":"Genoyer","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonathan","family":"Wilson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joshua M.","family":"Ames","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Caleb","family":"Stokes","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dante","family":"Moreno","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Noa","family":"Etzyon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew","family":"Oberst","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"suffix":"Jr.","given":"Michael","family":"Gale","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024061204250475000_2024.06.07.597966v1.1","doi-asserted-by":"crossref","first-page":"796","DOI":"10.1038\/s41564-020-0714-0","article-title":"The continued threat of emerging flaviviruses","volume":"5","year":"2020","journal-title":"Nat Microbiol"},{"key":"2024061204250475000_2024.06.07.597966v1.2","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.15585\/mmwr.mm7032a1","article-title":"West Nile Virus and Other Domestic Nationally Notifiable Arboviral Diseases - United States, 2019","volume":"70","year":"2021","journal-title":"MMWR Morb Mortal Wkly Rep"},{"key":"2024061204250475000_2024.06.07.597966v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2950"},{"key":"2024061204250475000_2024.06.07.597966v1.4","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01488-13"},{"key":"2024061204250475000_2024.06.07.597966v1.5","doi-asserted-by":"crossref","first-page":"e1009009","DOI":"10.1371\/journal.ppat.1009009","article-title":"MAVS regulates the quality of the antibody response to West-Nile Virus","volume":"16","year":"2020","journal-title":"PLoS Pathog"},{"key":"2024061204250475000_2024.06.07.597966v1.6","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000757"},{"key":"2024061204250475000_2024.06.07.597966v1.7","doi-asserted-by":"crossref","first-page":"3649","DOI":"10.1038\/s41467-019-11250-5","article-title":"RIG-I-like receptors direct inflammatory macrophage polarization against West Nile virus infection","volume":"10","year":"2019","journal-title":"Nat Commun"},{"key":"2024061204250475000_2024.06.07.597966v1.8","first-page":"110","article-title":"RIG-I in RNA virus recognition","volume":"479","year":"2015","journal-title":"Virology"},{"key":"2024061204250475000_2024.06.07.597966v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2011.05.003"},{"key":"2024061204250475000_2024.06.07.597966v1.10","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00814-06"},{"key":"2024061204250475000_2024.06.07.597966v1.11","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1515152113"},{"key":"2024061204250475000_2024.06.07.597966v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2015.06.015"},{"key":"2024061204250475000_2024.06.07.597966v1.13","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.83125-0"},{"key":"2024061204250475000_2024.06.07.597966v1.14","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00986-10"},{"key":"2024061204250475000_2024.06.07.597966v1.15","doi-asserted-by":"crossref","first-page":"4939","DOI":"10.1007\/s00018-021-03834-6","article-title":"Compartmentalized replication organelle of flavivirus at the ER and the factors involved","volume":"78","year":"2021","journal-title":"Cell. 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