Discovery of the Widespread Site-Specific Single-Stranded Nuclease Family Ssn

Discovery of the Widespread Site-Specific Single-Stranded Nuclease Family Ssn | 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 Article Discovery of the Widespread Site-Specific Single-Stranded Nuclease Family Ssn Frédéric Veyrier, Martin Chenal, Alex Rivera-Millot, Luke Harrisson, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4622751/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 10 Mar, 2025 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Site-specific endonucleases that exclusively cut single-stranded DNA have hitherto never been described and constitute a barrier to the development of ssDNA-based technologies. Here, we identify and characterize one such novel family of widely distributed site-specific single-stranded nucleases (Ssn) exhibiting unique ssDNA cleavage properties. This family is a member of the GIY-YIG nuclease superfamily that encompasses restriction enzymes, homing endonucleases, and transposases involved in important biological processes. Unlike other families of the GIY-YIG superfamily that typically possess additional functional domains that dictate their various specificities, Ssn are small proteins only consisting of the GIY-YIG domain. By first comprehensively studying the Ssn homolog from Neisseria meningitidis, we demonstrate that it interacts specifically with a sequence element (called NTS) present in hundreds of copies and surrounding important genes in pathogenic Neisseria. In this species, NTS/Ssn interactions modulate horizontal gene transfers through natural transformation and thus constitute a novel mechanism shaping genome dynamics. We further identified thousands of Ssn homologs throughout the bacterial kingdom and demonstrate, in vitro, a range of Ssn nuclease specificities for their corresponding NTS. We demonstrate proofs of concept for applications including ssDNA detection and digestion of ssDNA from Rolling Circle Amplification. This discovery and its applications set the stage for the development of innovative ssDNA-based molecular tools and technologies. Biological sciences/Biological techniques/Genetic engineering Biological sciences/Molecular biology/DNA recombination Biological sciences/Biotechnology/Molecular engineering/Synthetic biology Full Text Additional Declarations There is NO Competing Interest. M.C. and F.J.V. are co-inventors on the international patent submission PCT/CA2022/051668 related to this work. Supplementary Files SupplementarytablesV3.xlsx Related Manuscript File SupplementaryMaterialV5.pdf Cite Share Download PDF Status: Published Journal Publication published 10 Mar, 2025 Read the published version in Nature Communications → 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-4622751","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":319432960,"identity":"baffc395-fb5c-45b6-aff5-8dfa64d1a849","order_by":0,"name":"Frédéric 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