{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,9]],"date-time":"2026-11-09T13:08:16Z","timestamp":1794229696060,"version":"4.9.0"},"reference-count":52,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2004,5,21]],"date-time":"2004-05-21T00:00:00Z","timestamp":1085097600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2004,7]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p> <jats:italic>Histoplasma capsulatum<\/jats:italic> is a fungal pathogen that causes respiratory and systemic disease by proliferating within macrophages. While much is known about histoplasmosis, only a single virulence factor has been defined, in part because of the inefficiency of <jats:italic>Histoplasma<\/jats:italic> reverse genetics. As an alternative to allelic replacement, we have developed a telomeric plasmid\u2010based system for silencing gene expression in <jats:italic>Histoplasma<\/jats:italic> by RNA interference (RNAi). Episomal expression of long RNAs that form stem\u2013loop structures triggered gene silencing. To test the effectiveness of RNAi in <jats:italic>Histoplasma<\/jats:italic>, we depleted expression of a <jats:italic>gfp<\/jats:italic> transgene as well as two endogenous genes, <jats:italic>ADE2<\/jats:italic> and <jats:italic>URA5<\/jats:italic>, and showed significant reductions in corresponding gene function. Silencing was target gene specific, stable during macrophage infection and reversible. We used RNAi targeting <jats:italic>AGS1<\/jats:italic> (encoding \u03b1\u2010(1,3)\u2010glucan synthase) to deplete levels of \u03b1\u2010(1,3)\u2010glucan, a cell wall polysaccharide. Loss of \u03b1\u2010(1,3)\u2010glucan by RNAi yielded phenotypes indistinguishable from an <jats:italic>AGS1<\/jats:italic> deletion: attenuation of the ability to kill macrophages and colonize murine lungs. This demonstrates for the first time that \u03b1\u2010(1,3)\u2010glucan is an important contributor to <jats:italic>Histoplasma<\/jats:italic> virulence.<\/jats:p>","DOI":"10.1111\/j.1365-2958.2004.04131.x","type":"journal-article","created":{"date-parts":[[2004,7,2]],"date-time":"2004-07-02T06:32:38Z","timestamp":1088749958000},"page":"153-165","source":"Crossref","is-referenced-by-count":233,"title":["RNA interference in <i>Histoplasma capsulatum<\/i> demonstrates a role for \u03b1\u2010(1,3)\u2010glucan in virulence"],"prefix":"10.1111","volume":"53","author":[{"given":"Chad A.","family":"Rappleye","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacquelyn T.","family":"Engle","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William E.","family":"Goldman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2004,5,21]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.65.7.2564-2569.1997"},{"key":"e_1_2_6_3_1","first-page":"234","article-title":"Disseminated histoplasmosis in AIDS. 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