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Therefore, underscoring the urgent need for eco\u2010friendly alternatives to combat insecticide resistance and the scarcity of effective vaccines. Entomopathogenic fungi present a sustainable alternative to chemical insecticides; however, their widespread application is limited by their relatively low virulence.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>Here, we investigated the immunological interactions between <jats:italic>Metarhizium anisopliae<\/jats:italic> and <jats:italic>Aedes albopictus<\/jats:italic>, demonstrating that fungal infection significantly up\u2010regulated immune\u2010related genes in the Toll and melanization pathways, thereby enhancing antifungal and antibacterial defenses at 48\u2009h post\u2010infection (hpi). Small RNA sequencing identified miR339\u20105p as a crucial modulator, targeting the immune genes <jats:italic>Gram\u2010Negative Binding Protein 1<\/jats:italic> (<jats:italic>GNBP1<\/jats:italic>) and <jats:italic>CLIP\u2010domain Serine Protease B15<\/jats:italic> (<jats:italic>CLIPB15<\/jats:italic>), which are critical for Toll and phenoloxidase (PO) pathway activation. The administration of a synthetic miR339\u20105p mimic increased fungal virulence, resulting in a higher mortality rate among adult mosquitoes and a significant increase in the mortality rate of mosquito larvae within 24\u2009hpi. <jats:italic>GNBP1<\/jats:italic> was found to regulate both Toll and PO pathways, while <jats:italic>CLIPB15<\/jats:italic> specifically modulated the PO system by cleaving prophenoloxidase (PPO).<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSION<\/jats:title><jats:p>This research highlights the potential of leveraging <jats:italic>Ae. albopictus<\/jats:italic>\u2010encoded miR339\u20105p through advanced genetic engineering techniques to bolster the efficacy of existing fungal\u2010based mosquito control strategies, providing a promising approach in the fight against mosquito\u2010borne diseases. \u00a9 2024 Society of Chemical Industry.<\/jats:p><\/jats:sec>","DOI":"10.1002\/ps.8572","type":"journal-article","created":{"date-parts":[[2024,12,4]],"date-time":"2024-12-04T02:34:41Z","timestamp":1733279681000},"page":"1727-1739","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Boosting the efficacy of fungal biocontrol: <scp>miRNA339<\/scp>\u20105p\u2010mediated mosquito immunity regulation"],"prefix":"10.1002","volume":"81","author":[{"given":"Ruonan","family":"Zhang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Green Pesticide, College of Plant Protection South China Agricultural University  Guangzhou China"},{"name":"Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, College of Life Sciences South China Normal University  Guangzhou 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