Cellular uptake of extracellular dsRNA is tissue-dependent in insects

Cellular uptake of extracellular dsRNA is tissue-dependent in insects | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Cellular uptake of extracellular dsRNA is tissue-dependent in insects Xuekai Shi, Yaoming Liu, Xiaojian Liu, Mureed Abbas, Austin Merchant, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7156897/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 13 You are reading this latest preprint version Abstract Background RNA interference (RNAi), a naturally occurring gene silencing mechanism found in almost all eukaryotic organisms, has proven to be an adaptable and powerful tool in therapeutics, bioengineering, and agriculture. Differential responses to RNAi, however, are a key limiting factor, in which cellular uptake of exogenous dsRNA in target organisms remains poorly understood. Results Here, to fill this knowledge gap, we integrated omics tools with phenotypic assays to characterize dsRNA uptake mechanisms across tissues in the migratory locust, Locusta migratoria (Orthoptera). Our findings clearly demonstrate that cellular uptake of dsRNA is tissue-dependent, involving multiple cell membrane receptors and pathways. In hemocytes, uptake is rapid and mediated by clathrin-mediated endocytosis and macropinocytosis. Epidermal cells utilize clathrin- and caveolin-mediated endocytosis, while midgut cells employ caveolin-mediated endocytosis and Sid-like channel transport. Comparatively, clathrin-mediated endocytosis appears to be the most conserved mechanism across insects, including the red flour beetle, Tribolium castaneum (Coleoptera), and the Asian corn borer, Ostrinia furnacalis (Lepidoptera). Conclusions Taken together, dsRNA enters the cells of different tissue types through diverse pathways. This systematic and comprehensive study not only advances our understanding of the cellular uptake of extracellular dsRNA and the resultant differential sensitivity to RNAi in insects, but also facilitates the ongoing integration of this species-specific biotechnology into sustainable Integrated Pest Management practices. cellular uptake dsRNA tissue-dependent Locusta migratoria endocytosis RNAi Full Text Additional Declarations No competing interests reported. Supplementary Files Shietal.SupplementaryInformation.doc Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 10 Oct, 2025 Reviews received at journal 09 Oct, 2025 Reviews received at journal 05 Oct, 2025 Reviews received at journal 03 Oct, 2025 Reviewers agreed at journal 27 Sep, 2025 Reviewers agreed at journal 26 Sep, 2025 Reviewers agreed at journal 25 Sep, 2025 Reviews received at journal 11 Aug, 2025 Reviewers agreed at journal 05 Aug, 2025 Reviewers invited by journal 05 Aug, 2025 Editor assigned by journal 22 Jul, 2025 Submission checks completed at journal 21 Jul, 2025 First submitted to journal 18 Jul, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7156897","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":495973458,"identity":"1bfcd501-8d0b-47c1-8bd7-c9c8d684f9db","order_by":0,"name":"Xuekai Shi","email":"","orcid":"","institution":"Shanxi University","correspondingAuthor":false,"prefix":"","firstName":"Xuekai","middleName":"","lastName":"Shi","suffix":""},{"id":495973459,"identity":"d15d1e73-92db-44d7-af7d-7c236c4fea61","order_by":1,"name":"Yaoming Liu","email":"","orcid":"","institution":"Shanxi 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Differential responses to RNAi, however, are a key limiting factor, in which cellular uptake of exogenous dsRNA in target organisms remains poorly understood.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eHere, to fill this knowledge gap, we integrated omics tools with phenotypic assays to characterize dsRNA uptake mechanisms across tissues in the migratory locust, \u003cem\u003eLocusta migratoria\u003c/em\u003e (Orthoptera). Our findings clearly demonstrate that cellular uptake of dsRNA is tissue-dependent, involving multiple cell membrane receptors and pathways. In hemocytes, uptake is rapid and mediated by clathrin-mediated endocytosis and macropinocytosis. Epidermal cells utilize clathrin- and caveolin-mediated endocytosis, while midgut cells employ caveolin-mediated endocytosis and Sid-like channel transport. Comparatively, clathrin-mediated endocytosis appears to be the most conserved mechanism across insects, including the red flour beetle, \u003cem\u003eTribolium castaneum\u003c/em\u003e (Coleoptera), and the Asian corn borer, \u003cem\u003eOstrinia furnacalis\u003c/em\u003e (Lepidoptera).\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e\u003cp\u003eTaken together, dsRNA enters the cells of different tissue types through diverse pathways. This systematic and comprehensive study not only advances our understanding of the cellular uptake of extracellular dsRNA and the resultant differential sensitivity to RNAi in insects, but also facilitates the ongoing integration of this species-specific biotechnology into sustainable Integrated Pest Management practices.\u003c/p\u003e","manuscriptTitle":"Cellular uptake of extracellular dsRNA is tissue-dependent in insects","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-08 20:07:47","doi":"10.21203/rs.3.rs-7156897/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-10-10T16:50:33+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-09T12:34:02+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-06T00:44:43+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-03T11:47:15+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"182947485808597965098575952835144881183","date":"2025-09-28T01:49:11+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"9913268286651172131534493003895934095","date":"2025-09-26T13:59:42+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"231375739425967550200637529554050106458","date":"2025-09-25T13:35:52+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-11T14:53:04+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"104165724614988653990862365011766453692","date":"2025-08-05T11:11:19+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-08-05T08:59:06+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-07-22T08:26:42+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-07-21T05:28:40+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Biology","date":"2025-07-18T10:39:29+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-biology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [BMC Biology](https://bmcbiol.biomedcentral.com/)","snPcode":"12915","submissionUrl":"https://submission.springernature.com/new-submission/12915/3","title":"BMC Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"36e848a6-55f2-4c38-a9cd-fbdf8ac12154","owner":[],"postedDate":"August 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-01-16T14:53:57+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-08 20:07:47","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7156897","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7156897","identity":"rs-7156897","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}