Copper Ion Genetic Biosensor: A Synthetic Biology Approach for Environmental Heavy Metal Detection

Copper Ion Genetic Biosensor: A Synthetic Biology Approach for Environmental Heavy Metal Detection | 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 Copper Ion Genetic Biosensor: A Synthetic Biology Approach for Environmental Heavy Metal Detection Yaman Yazici This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7066326/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 Excess environmental copper from industrial and agricultural sources poses significant risks to ecosystems and human health. We engineered a whole-cell Escherichia coli biosensor to rapidly detect bioavailable Cu(II) ions using a copper-inducible genetic circuit. The system couples the E. coli copA promoter – a copper- responsive regulatory element – to a green fluorescent protein (GFP) reporter gene, producing a quantifiable fluorescence signal in the presence of copper. We constructed the sensor on a plasmid and transformed it into E. coli ; subsequent assays measured GFP fluorescence across a gradient of copper concentrations. The biosensor responded to Cu(II) in a dose-dependent manner, with a detection threshold of ~ 10 µM and a linear dynamic range up to ~ 100 µM. It demonstrated specificity for copper over other common metal ions, and maintained performance in spiked environmental water samples. Time-course experiments further showed that detectable fluorescence could be induced within hours of exposure. These results underscore the potential of synthetic biology for creating cost-effective, field-deployable heavy metal detectors. Our engineered copper biosensor offers real-time, in situ monitoring capability and could be expanded or optimized for improved sensitivity and broader heavy-metal detection. We discuss its strengths and limitations relative to existing methods, and propose future enhancements – such as circuit modifications and cellular engineering – to meet stringent environmental regulatory standards. This work provides a proof- of-concept for deploying genetically engineered whole-cell sensors as practical tools for environmental monitoring of heavy metal contamination. Full Text Additional Declarations No competing interests reported. Supplementary Files NoGelsorBlotsSupplementaryFile.docx 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-7066326","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":489747383,"identity":"b2cc62f0-13f5-49b1-b217-ef08c9fac926","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":"2025-07-07 14:08:37","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7066326/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7066326/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":88129167,"identity":"2e92cc37-4a2f-49f0-8636-8b98d3b9b6b3","added_by":"auto","created_at":"2025-08-01 18:16:22","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":722764,"visible":true,"origin":"","legend":"","description":"","filename":"CopperIonGeneticBiosensorASyntheticBiologyApproachforEnvironmentalHeavyMetalDetection.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7066326/v1_covered_e444e9da-5630-4520-9367-b5d16749be97.pdf"},{"id":87590465,"identity":"cb581a04-6fa3-40de-bd7d-dee61d3dc473","added_by":"auto","created_at":"2025-07-25 14:42:52","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":13554,"visible":true,"origin":"","legend":"","description":"","filename":"NoGelsorBlotsSupplementaryFile.docx","url":"https://assets-eu.researchsquare.com/files/rs-7066326/v1/6f9b70102099dbbdcd6dcd2a.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Copper Ion Genetic Biosensor: A Synthetic Biology Approach for Environmental Heavy Metal Detection","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-7066326/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7066326/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eExcess environmental copper from industrial and agricultural sources poses significant risks to ecosystems and human health. We engineered a whole-cell \u003cem\u003eEscherichia coli\u003c/em\u003e biosensor to rapidly detect bioavailable Cu(II) ions using a copper-inducible genetic circuit. The system couples the \u003cem\u003eE. coli\u003c/em\u003e copA promoter – a copper- responsive regulatory element – to a green fluorescent protein (GFP) reporter gene, producing a quantifiable fluorescence signal in the presence of copper. We constructed the sensor on a plasmid and transformed it into \u003cem\u003eE. coli\u003c/em\u003e; subsequent assays measured GFP fluorescence across a gradient of copper concentrations. The biosensor responded to Cu(II) in a dose-dependent manner, with a detection threshold of ~ 10 µM and a linear dynamic range up to ~ 100 µM. It demonstrated specificity for copper over other common metal ions, and maintained performance in spiked environmental water samples. Time-course experiments further showed that detectable fluorescence could be induced within hours of exposure. 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