OptoRibo-seq for spatiotemporally resolved mapping of local protein translatome

OptoRibo-seq for spatiotemporally resolved mapping of local protein translatome | 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 OptoRibo-seq for spatiotemporally resolved mapping of local protein translatome Peng Zou, Ruixuan Wang, Ruixiang Wang, Wentao Wang, Yanjun Liu, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3982122/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 Localized protein translation occurs in numerous subcellular compartments and regulates diverse biological processes by rapidly changing protein compositions in response to subcellular needs. Existing assays for subcellular local translation either require physical isolation, which is prone to contamination and loss of material, or imaging-based readout, which is often hampered with low throughput. In this study, we report the development of optoRibo-seq method that features photoactivatable enzyme-mediated proximity labeling of ribosomes in genetically specified subcellular locations. We demonstrate the spatial specificity of optoRibo-seq at the endoplasmic reticulum (ER) membrane and mitochondrial outer membrane. OptoRibo-seq is further applied to map the dynamic changes in the ER-proximal translatome in cells undergoing chemically induced ER stress, identifying transcripts involved in protein folding and amino acid transport. Our strategy provides a general platform for spatiotemporally resolved profiling of subcellular protein translation in various organelles. Biological sciences/Chemical biology/RNA Biological sciences/Chemical biology/Chemical tools Proximity labeling Genetic codon expansion Ribosome profiling Biotin ligase ER stress Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryData1.xlsx Dataset 1 SupplementaryData2A.xlsx Dataset 2A SupplementaryData2B.xlsx Dataset 2B SupplementaryData3.xlsx Dataset 3 SupplementaryData4A.xlsx Dataset 4A SupplementaryData4B.xlsx Dataset 4B Supplementaryinformationv4.0.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. 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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-3982122","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":275760921,"identity":"5a3e6b57-4933-4032-85d2-151528131a28","order_by":0,"name":"Peng 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