Resurrection of chromosomes from frozen animals by single chromosome transfer into mouse oocytes

Resurrection of chromosomes from frozen animals by single chromosome transfer into mouse oocytes | 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 Resurrection of chromosomes from frozen animals by single chromosome transfer into mouse oocytes Sayaka Wakayama, Ryoko Araki, Misato Sunayama, Kazuo Yamagata, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9131912/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Reviving extinct animals offers a crucial opportunity to recover lost or unknown genetic resources, yet cloning methods are unsuitable because they depend on intact donor nuclei and abundant oocytes or recipients from closely related species. To overcome these constraints, we explored a chromosome level revival strategy. Blood cells from rat carcasses stored at − 30°C for over one year were introduced into enucleated mouse oocytes, where the rat nuclei underwent premature chromosome condensation. Microtubule polymerization inhibition enabled dispersion of rat chromosomes within the ooplasm, allowing isolation of individual chromosomes by micromanipulation. Each chromosome was subsequently transferred into an intact mouse oocyte, followed by intracytoplasmic sperm injection using GFP-transgenic mouse sperm. Embryos were cultured to the blastocyst stage, yielding 17 ES cell lines, two of which carried 41 chromosomes. Spectral karyotyping confirmed the presence of rat chromosome 9 alongside a full set of normal mouse chromosomes. These ES cells generated chimeric mice exhibiting GFP based chimerism across multiple organs. Histological analyses further demonstrated expression of numerous genes located on rat chromosome 9 within chimera mouse. This study demonstrated that a single chromosome from a frozen extinct species can be functionally revived and its transcriptional activity assessed within an interspecies oocyte. Biological sciences/Biological techniques Biological sciences/Cell biology Biological sciences/Developmental biology Biological sciences/Genetics Interspecies chromosome transfer clone resurrection extinct species Full Text Additional Declarations No competing interests reported. Supplementary Files Supplementarydata1PredictionofratHiCEPfragments.xlsx Supplementarymovie1.mp4 Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 29 Apr, 2026 Reviews received at journal 18 Apr, 2026 Reviews received at journal 09 Apr, 2026 Reviewers agreed at journal 04 Apr, 2026 Reviewers agreed at journal 03 Apr, 2026 Reviewers invited by journal 26 Mar, 2026 Editor assigned by journal 26 Mar, 2026 Editor invited by journal 26 Mar, 2026 Submission checks completed at journal 25 Mar, 2026 First submitted to journal 25 Mar, 2026 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-9131912","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":613484668,"identity":"8e00c3fa-68c6-4141-bf92-129dccb5343e","order_by":0,"name":"Sayaka Wakayama","email":"","orcid":"","institution":"University of Yamanashi","correspondingAuthor":false,"prefix":"","firstName":"Sayaka","middleName":"","lastName":"Wakayama","suffix":""},{"id":613484669,"identity":"88b122da-598a-4f3b-be6a-f2e480fbcedc","order_by":1,"name":"Ryoko Araki","email":"","orcid":"","institution":"National Institutes for Quantum Science and 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