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However, the mechanisms underlying <jats:italic>Wolbachia<\/jats:italic>-mediated virus blockade have not been fully elucidated. Here, we report that <jats:italic>Wolbachia<\/jats:italic> activates the host cytoplasmic miRNA biogenesis pathway to suppress DENV infection. Through the suppression of the long noncoding RNA aae-lnc-2268 by <jats:italic>Wolbachia w<\/jats:italic>AlbB, aae-miR-34-3p, a miRNA upregulated by the <jats:italic>Wolbachia<\/jats:italic> strains <jats:italic>w<\/jats:italic>AlbB and <jats:italic>w<\/jats:italic>MelPop, promoted the expression of the antiviral effector <jats:italic>defensin<\/jats:italic> and <jats:italic>cecropin<\/jats:italic> genes through the Toll pathway regulator <jats:italic>MyD88<\/jats:italic>. Notably, anti-DENV resistance induced by <jats:italic>Wolbachia<\/jats:italic> can be further enhanced, with the potential to achieve complete virus blockade by increasing the expression of aae-miR-34-3p in <jats:italic>Ae<\/jats:italic>. <jats:italic>aegypti<\/jats:italic>. Furthermore, the downregulation of aae-miR-34-3p compromised <jats:italic>Wolbachia<\/jats:italic>-mediated virus blockade. These findings reveal a novel mechanism by which <jats:italic>Wolbachia<\/jats:italic> establishes crosstalk between the cytoplasmic miRNA pathway and the Toll pathway via aae-miR-34-3p to strengthen antiviral immune responses against DENV. Our results will aid in the advancement of <jats:italic>Wolbachia<\/jats:italic> for arbovirus control by enhancing its virus-blocking efficiency.<\/jats:p>","DOI":"10.1371\/journal.ppat.1012296","type":"journal-article","created":{"date-parts":[[2024,6,17]],"date-time":"2024-06-17T18:13:30Z","timestamp":1718648010000},"page":"e1012296","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":21,"title":["Wolbachia mediates crosstalk between miRNA and Toll pathways to enhance resistance to dengue virus in Aedes aegypti"],"prefix":"10.1371","volume":"20","author":[{"given":"Lingzhi","family":"She","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengyi","family":"Shi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting","family":"Cao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Yuan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renke","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weifeng","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yueting","family":"She","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chaojun","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qin","family":"Zeng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Mao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yalan","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7786-012X","authenticated-orcid":true,"given":"Zhiyong","family":"Xi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4726-5095","authenticated-orcid":true,"given":"Xiaoling","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,6,17]]},"reference":[{"issue":"6","key":"ppat.1012296.ref001","doi-asserted-by":"crossref","first-page":"e2021055522","DOI":"10.1542\/peds.2021-055522","article-title":"Dengue: a growing problem with new interventions","volume":"149","author":"JM Wong","year":"2022","journal-title":"Pediatrics"},{"issue":"12","key":"ppat.1012296.ref002","doi-asserted-by":"crossref","first-page":"1495","DOI":"10.1080\/14787210.2021.1949983","article-title":"Dengue vaccine: an update","volume":"19","author":"CH Huang","year":"2021","journal-title":"Expert Rev Anti Infect Ther"},{"issue":"5746","key":"ppat.1012296.ref003","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1126\/science.1117607","article-title":"Wolbachia establishment and invasion in an Aedes aegypti laboratory population","volume":"310","author":"Z Xi","year":"2005","journal-title":"Science"},{"issue":"7767","key":"ppat.1012296.ref004","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1038\/s41586-019-1407-9","article-title":"Incompatible and sterile insect techniques combined eliminate mosquitoes","volume":"572","author":"X Zheng","year":"2019","journal-title":"Nature"},{"issue":"7","key":"ppat.1012296.ref005","doi-asserted-by":"crossref","first-page":"e1754","DOI":"10.1371\/journal.pntd.0001754","article-title":"Wolbachia induces density-dependent inhibition to dengue virus in mosquito cells.","volume":"6","author":"P Lu","year":"2012","journal-title":"PLoS Negl Trop Dis"},{"issue":"7","key":"ppat.1012296.ref006","doi-asserted-by":"crossref","first-page":"1268","DOI":"10.1016\/j.cell.2009.11.042","article-title":"A Wolbachia symbiont in Aedes aegypti limits infection with dengue, chikungunya, and Plasmodium","volume":"139","author":"LA Moreira","year":"2009","journal-title":"Cell"},{"issue":"6133","key":"ppat.1012296.ref007","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1126\/science.1236192","article-title":"Wolbachia invades Anopheles stephensi populations and induces refractoriness to Plasmodium infection","volume":"340","author":"G Bian","year":"2013","journal-title":"Science"},{"issue":"1","key":"ppat.1012296.ref008","first-page":"E23","article-title":"Wolbachia induces reactive oxygen species (ROS)-dependent activation of the Toll pathway to control dengue virus in the mosquito Aedes aegypti.","volume":"109","author":"X Pan","year":"2012","journal-title":"Proc. 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