Proteus Project - CRISPR-Based Programmable Therapeutics for Targeted Apoptosis in Cancer Cells

Proteus Project - CRISPR-Based Programmable Therapeutics for Targeted Apoptosis in Cancer Cells | 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 Proteus Project - CRISPR-Based Programmable Therapeutics for Targeted Apoptosis in Cancer Cells Yaman Yazici This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7160959/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Cancer therapy faces a critical need for treatments that selectively eliminate malignant cells without harming healthy tissue. The Proteus Project addresses this challenge by engineering a programmable gene circuit that couples CRISPR-based RNA sensing with an inducible cell death mechanism. Specifically, we repurpose a novel Type III-E CRISPR-Cas system (“Craspase”) – an RNA-guided protease complex – to detect cancer-specific RNA transcripts and, in response, cleave engineered gasdermin fusion proteins to trigger cell death (1)(2). We constructed the Proteus system in Saccharomyces cerevisiae as a surrogate model, integrating components that sense an oncogenic KRAS mutation and execute targeted pyroptosis (inflammatory apoptosis-like cell death). Preliminary results demonstrate successful assembly of the Craspase– gasdermin circuit, expression of key proteins, and proof-of-concept cell killing specifically in the presence of the oncogenic RNA trigger. This work, conducted as part of the iGEM 2025 SynBio Collective, showcases a modular synthetic biology approach for precision oncology therapeutics. The ongoing study underscores the potential of CRISPR-guided proteases in in situ cancer cell ablation and sets the stage for future validation in mammalian systems. Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementaryFileProteus.pdf Cite Share Download PDF Status: Posted Version 1 posted 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-7160959","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":491001142,"identity":"4b2d7423-6d9b-4ff6-b146-32a3810fe376","order_by":0,"name":"Yaman Yazici","email":"data:image/png;base64,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","orcid":"","institution":"McGill University","correspondingAuthor":true,"prefix":"","firstName":"Yaman","middleName":"","lastName":"Yazici","suffix":""}],"badges":[],"createdAt":"2025-07-18 23:53:07","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7160959/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7160959/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":90350844,"identity":"5518411e-6f79-4e2f-ac3c-af692b1c5852","added_by":"auto","created_at":"2025-09-01 17:46:33","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":614018,"visible":true,"origin":"","legend":"","description":"","filename":"ProteusProjectCRISPRBasedProgrammableTherapeuticsforTargetedApoptosisinCancerCells.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7160959/v1_covered_bc20108f-211e-4c65-b630-2ce896917e74.pdf"},{"id":87795334,"identity":"048d604b-fd81-437a-9203-e0ab5d31b5d4","added_by":"auto","created_at":"2025-07-29 06:50:37","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":661894,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryFileProteus.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7160959/v1/99425f41ecf8569028e506cf.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Proteus Project - CRISPR-Based Programmable Therapeutics for Targeted Apoptosis in Cancer Cells","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-7160959/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7160959/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCancer therapy faces a critical need for treatments that selectively eliminate malignant cells without harming healthy tissue. 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