Lentivirus Enables the Detection of Strand-Specific ssDNA Gaps by the DNA Fiber Spreading Assay

Lentivirus Enables the Detection of Strand-Specific ssDNA Gaps by the DNA Fiber Spreading Assay | 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 Lentivirus Enables the Detection of Strand-Specific ssDNA Gaps by the DNA Fiber Spreading Assay Jieya shao, Melisande Wong, Xupei Ou, Evan Brown Ton This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8265086/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 The single-molecule DNA fiber spreading assay is widely used to study DNA replication fork dynamics through pulse labeling of nascent DNA with distinct thymidine analogs. However, tight sister chromatid cohesion during spreading causes leading and lagging strands to appear as a single fiber, masking strand-specific replication changes that are increasingly recognized as biologically and therapeutically important. Here, we report the unexpected finding that lentiviral infection of human cell lines prior to DNA spreading enables visualization of lagging strand–specific single-stranded DNA gaps arising from defects in Okazaki fragment maturation. Our results suggest that such an effect is transient and independent of viral vectors, encoded sequences, or genome integration. Mechanistically, lentivirus reduces cohesin level on nascent DNA, indicating that transient loosening of sister chromatid cohesion allows daughter strand separation. We propose that pre-exposure to control lentiviruses can be a simple, effective modification to enable the DNA fiber spreading assay to detect strand-specific DNA replication changes and overcome its limitation. Biological sciences/Molecular biology/DNA replication Biological sciences/Biological techniques 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. 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-8265086","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":559193079,"identity":"f3d5157f-6248-4cb7-9f15-c5988edfbe00","order_by":0,"name":"Jieya 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