{"paper_id":"2cbeae4f-65f6-4c0a-9974-d99c6c3a7e64","body_text":"PRINTS, a tool for subcellular protein and organelle translocation and its application in screening LLPS disrupting peptides | 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 PRINTS, a tool for subcellular protein and organelle translocation and its application in screening LLPS disrupting peptides Hanzeng Li, Cheng Tang, Qian Zhou, Ziqing Gao, Pei Peng, Jinsen He, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7873023/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Protein subcellular localization is dynamically regulated, requiring new tools to control protein translocation for elucidating its biological significance and eventually enabling synthetic biology applications. Here, we developed a new method, Protein Rerouting via INtein-mediated Trans-Splicing (PRINTS), for precise control of protein translocation across diverse subcellular compartments. By reconstituting functional signaling peptides, PRINTS enables robust relocalization to varied destinations, including the 26S proteasome, nucleus, mitochondria, plasma membrane, endomembrane organelles, and liquid-liquid phase separation (LLPS) condensates. Moreover, integration of the CRY2clust light-responsive dimerization domain into PRINTS achieved optically controlled protein entry into the cell membrane and LLPS condensates. Additionally, PRINTS allow light-controlled organelle relocalization, such as LLPS condensates or mitochondria to the plasma membrane. We also applied this targeted delivery system to screen and validate de novo designed short peptides that disrupt LLPS. As a versatile platform, PRINTS facilitate precise manipulation of protein and organelle dynamics, offering a powerful tool to study the regulatory coordination between membranous and membraneless compartments in response to physiological needs. Biological sciences/Cell biology/Organelles Biological sciences/Biological techniques/Molecular engineering/Synthetic biology Full Text Additional Declarations There is NO Competing Interest. Cite Share Download PDF Status: Under Review 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. 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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-7873023\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Article\",\"associatedPublications\":[],\"authors\":[{\"id\":538859646,\"identity\":\"20d2d5c0-86c8-482f-b349-2668d46bfe81\",\"order_by\":0,\"name\":\"Hanzeng 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