Development of B-to-A base editors by leveraging translesion synthesis polymerase

Development of B-to-A base editors by leveraging translesion synthesis polymerase | 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 Development of B-to-A base editors by leveraging translesion synthesis polymerase Puping Liang, Pengfei Zhang, Yuxi Chen, Weijun Zhao, Yunqian Zhang, and 20 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7122028/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 Base editing technology enables single-nucleotide conversions without donor templates or DNA double-strand breaks (DSBs), yet current base editors lack the capacity to efficiently address C-to-A, G-to-A, or T-to-A conversion. Here we report the development of B-to-A base editors (BABEs, B=C/G/T) through co-expressing certain glycosylase-based base editors together with the engineered translesion synthesis (TLS) polymerase polη Q38S variant, achieving about an average of 5-fold increase in B-to-A editing purity and efficiency. The C-to-A base editor (CABE), G-to-A base editor (GABE), and T-to-A base editor (TABE) achieved C-to-A purity up to 69.0%, G-to-A purity up to 57.1% and T-to-A purity up to 91.6%, respectively, in human cells without compromising the editing efficiency. In addition, we enhanced the editing efficiency of gGBE through rational design and further developed more efficient G-to-A base editor (GABEv2), which displayed a 2.1-fold improvement in G-to-A editing purity and efficiency compared to gGBE in mouse embryo. Our study expands current base editor toolbox by providing B-to-A base editors, broadening the applications of base editors in fundamental research and clinical therapeutics. Biological sciences/Molecular biology/DNA damage and repair Biological sciences/Systems biology/Molecular engineering Biological sciences/Biological techniques/Genetic engineering Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryTable1.pdf Supplementary Table 1 SupplementaryTable2.pdf Supplementary Table 2 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. 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-7122028","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":490213174,"identity":"fe102594-b156-4386-8188-df397ec1c7de","order_by":0,"name":"Puping 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