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RNA interference (RNAi) in C. chilonis is challenging due to the unique biological traits of endoparasitoids. In this study, dsRNA was delivered to C. chilonis by microinjection of pupae within cocoons (delivery outside the host) and nanomaterial-mediated immersion, which delivered dsRNA to larvae inside the host. Both systems were effective in silencing the target gene, Cccaspase-1, with microinjection achieving 41.34% RNAi efficiency in 4-d-old pupae, and nanomaterial immersion exhibiting a slightly higher efficacy (45.63%); furthermore, the results indicated a crucial role for Cccaspase-1 in the growth and development of C. chilonis. Delivery of dsRNA by microinjection offers a high level of precision and is suitable for small-scale genetic screening, whereas nanomaterial immersion enables high-throughput delivery of dsRNA and can be used to monitor silencing effects throughout the development of C. chilonis. This study provides key insights for optimizing RNAi techniques in endoparasitoid wasps.<\/jats:p>","DOI":"10.1093\/jee\/toag299","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T11:48:42Z","timestamp":1789127322000},"source":"Crossref","is-referenced-by-count":0,"title":["Functional analysis of\n                    <i>Caspase-1<\/i>\n                    in\n                    <i>Cotesia chilonis<\/i>\n                    via pupal microinjection and larval nanomaterial immersion-mediated RNA interference"],"prefix":"10.1093","author":[{"given":"Qi-Yan","family":"He","sequence":"first","affiliation":[{"name":"College of Plant Protection, Institute of Applied Entomology, Yangzhou University , Yangzhou,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fu-Jing","family":"He","sequence":"additional","affiliation":[{"name":"School of Tourism & Culinary Arts, Wuxi Vocational Institute of Commerce , Wuxi,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Wan","sequence":"additional","affiliation":[{"name":"College of Plant Protection, Institute of Applied Entomology, Yangzhou University , Yangzhou,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong-Yu","family":"Qian","sequence":"additional","affiliation":[{"name":"College of Plant Protection, Institute of Applied Entomology, Yangzhou University , Yangzhou,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian-Fei","family":"Mei","sequence":"additional","affiliation":[{"name":"Guizhou Zhuohao Agricultural Science and Technology Co., Ltd. , Zunyi,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8243-1423","authenticated-orcid":false,"given":"Yu-Zhou","family":"Du","sequence":"additional","affiliation":[{"name":"College of Plant Protection, Institute of Applied Entomology, Yangzhou University , Yangzhou,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0009-2761","authenticated-orcid":false,"given":"Ming-Xing","family":"Lu","sequence":"additional","affiliation":[{"name":"College of Plant Protection, Institute of Applied Entomology, Yangzhou University , Yangzhou,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,9,29]]},"reference":[{"key":"2026092912184705700_toag299-B1","doi-asserted-by":"publisher","first-page":"211","DOI":"10.3390\/insects11040211","article-title":"Male-biased adult production of the striped fruit fly, Zeugodacus scutellata, by feeding dsRNA specific to Transformer-2","volume":"11","author":"Al Baki","year":"2020","journal-title":"Insects"},{"key":"2026092912184705700_toag299-B2","doi-asserted-by":"publisher","first-page":"1219","DOI":"10.1002\/PS.7851","article-title":"CRISPR\/Cas9-mediated mutagenesis of the white gene in an ectoparasitic wasp, Habrobracon hebetor","volume":"80","author":"Bai","year":"2024","journal-title":"Pest Manag. 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