{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,18]],"date-time":"2026-06-18T22:32:40Z","timestamp":1781821960219,"version":"3.54.5"},"reference-count":44,"publisher":"National Academy of Sciences","issue":"49","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2004,12,7]]},"abstract":"<jats:p>\n                    RNA interference (RNAi) is triggered in eukaryotic organisms by double-stranded RNA (dsRNA), and it destroys any mRNA that has sequence identity with the dsRNA trigger. The RNAi pathway in\n                    <jats:italic>Anopheles gambiae<\/jats:italic>\n                    can be silenced by transfecting cells with dsRNA derived from exon sequence of the\n                    <jats:italic>A. gambiae Argonaute2<\/jats:italic>\n                    (\n                    <jats:italic>AgAgo2<\/jats:italic>\n                    ) gene. We hypothesized that RNAi may also act as an antagonist to alphavirus replication in\n                    <jats:italic>A. gambiae<\/jats:italic>\n                    because RNA viruses form dsRNA during replication. Silencing\n                    <jats:italic>AgAgo2<\/jats:italic>\n                    expression would make\n                    <jats:italic>A. gambiae<\/jats:italic>\n                    mosquitoes more permissive to virus infection. To determine whether RNAi conditions the vector competence of\n                    <jats:italic>A. gambiae<\/jats:italic>\n                    for O'nyong-nyong virus (ONNV), we engineered a genetically modified ONNV that expresses enhanced GFP (eGFP) as a marker. After intrathoracic injection, ONNV-eGFP slowly spread to other\n                    <jats:italic>A. gambiae<\/jats:italic>\n                    tissues over a 9-day incubation period. Mosquitoes were then coinjected with virus and either control \u03b2-\n                    <jats:italic>galactosidase<\/jats:italic>\n                    dsRNA (ds\u03b2gal; note that \u201cds\u201d is used as a prefix to indicate the dsRNA derived from a given gene throughout) or ONNV dsnsP3. Treatment with dsnsP3 inhibited virus spread significantly, as determined by eGFP expression patterns. ONNV-eGFP titers from mosquitoes coinjected with dsnsP3 were significantly lower at 3 and 6 days after injection than in mosquitoes coinjected with ds\u03b2gal. Mosquitoes were then coinjected with ONNV-eGFP and dsAgAgo2. Mosquitoes coinjected with virus and\n                    <jats:italic>AgAgo2<\/jats:italic>\n                    dsRNA displayed widespread eGFP expression and virus titers 16-fold higher than ds\u03b2gal controls after 3 or 6 days after injection. These observations provide direct evidence that RNAi is an antagonist of ONNV replication in\n                    <jats:italic>A. gambiae<\/jats:italic>\n                    , and they suggest that the innate immune response conditions vector competence.\n                  <\/jats:p>","DOI":"10.1073\/pnas.0406983101","type":"journal-article","created":{"date-parts":[[2004,12,6]],"date-time":"2004-12-06T20:24:54Z","timestamp":1102364694000},"page":"17240-17245","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":289,"title":["RNA interference acts as a natural antiviral response to O'nyong-nyong virus (\n                    <i>Alphavirus<\/i>\n                    ; Togaviridae) infection of\n                    <i>Anopheles gambiae<\/i>"],"prefix":"10.1073","volume":"101","author":[{"given":"Kimberly M.","family":"Keene","sequence":"first","affiliation":[{"name":"Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian D.","family":"Foy","sequence":"additional","affiliation":[{"name":"Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irma","family":"Sanchez-Vargas","sequence":"additional","affiliation":[{"name":"Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barry J.","family":"Beaty","sequence":"additional","affiliation":[{"name":"Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carol D.","family":"Blair","sequence":"additional","affiliation":[{"name":"Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ken E.","family":"Olson","sequence":"additional","affiliation":[{"name":"Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 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