{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T06:29:32Z","timestamp":1791872972936,"version":"4.3.4"},"reference-count":49,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2024,12,16]],"date-time":"2024-12-16T00:00:00Z","timestamp":1734307200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01AI150478"],"award-info":[{"award-number":["R01AI150478"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01GM118474"],"award-info":[{"award-number":["R01GM118474"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000862","name":"Doris Duke Foundation","doi-asserted-by":"crossref","award":["DDF #2022092"],"award-info":[{"award-number":["DDF #2022092"]}],"id":[{"id":"10.13039\/100000862","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100007920","name":"University of Massachusetts Chan Medical School","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007920","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,1,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Recently, cytosine base editors (CBEs) have emerged as a promising therapeutic tool for specific editing of single nucleotide variants and disrupting specific genes associated with disease. Despite this promise, the currently available CBEs have the significant liabilities of off-target and bystander editing activities, partly due to the mechanism by which they are delivered, causing limitations in their potential applications. In this study, we engineered optimized, soluble and stable Cas-embedded CBEs (CE_CBEs) that integrate several recent advances, which were efficiently formulated for direct delivery into cells as ribonucleoprotein (RNP) complexes. Our resulting CE_CBE RNP complexes efficiently target cytosines in TC dinucleotides with minimal off-target or bystander mutations. Delivery of additional uracil glycosylase inhibitor protein in trans further increased C-to-T editing efficiency and target purity in a dose-dependent manner, minimizing indel formation. A single electroporation was sufficient to effectively edit the therapeutically relevant locus BCL11A for sickle cell disease in hematopoietic stem and progenitor cells in a dose-dependent manner without cellular toxicity. Significantly, these CE_CBE RNPs permitted highly efficient editing and engraftment of transplanted cells in mice. Thus, our designed CBE proteins provide promising reagents for RNP-based editing at disease-related sites.<\/jats:p>","DOI":"10.1093\/nar\/gkae1217","type":"journal-article","created":{"date-parts":[[2024,12,16]],"date-time":"2024-12-16T10:07:56Z","timestamp":1734343676000},"source":"Crossref","is-referenced-by-count":9,"title":["Direct delivery of Cas-embedded cytosine base editors as ribonucleoprotein complexes for efficient and accurate editing of clinically relevant targets"],"prefix":"10.1093","volume":"53","author":[{"given":"Jeong Min","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School , 364 Plantation Street,\u00a0 Worcester , MA\u00a0 01605 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Zeng","sequence":"additional","affiliation":[{"name":"Division of Hematology\/Oncology, Boston Children\u2019s Hospital , 1 Blackfan Circle,\u00a0Boston, MA 02115 ,","place":["USA"]},{"name":"Department of Pediatric Oncology, Dana-Farber Cancer Institute , 450 Brookline Avenue,\u00a0Boston, MA 02215 ,","place":["USA"]},{"name":"Department of Pediatrics, Harvard Stem Cell Institute, Broad Institute of Harvard and MIT, Harvard Medical School , 25 Shattuck Street, Boston, MA 02115 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengpeng","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Molecular, Cell and Cancer Biology, UMass Chan Medical School , 364 Plantation Street,\u00a0Worcester, MA 01605 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"My Anh","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Division of Hematology\/Oncology, Boston Children\u2019s Hospital , 1 Blackfan Circle,\u00a0Boston, MA 02115 ,","place":["USA"]},{"name":"Department of Pediatric Oncology, Dana-Farber Cancer Institute , 450 Brookline Avenue,\u00a0Boston, MA 02215 ,","place":["USA"]},{"name":"Department of Pediatrics, Harvard Stem Cell Institute, Broad Institute of Harvard and MIT, Harvard Medical School , 25 Shattuck Street, Boston, MA 02115 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diego","family":"Suchenski\u00a0Loustaunau","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School , 364 Plantation Street,\u00a0 Worcester , MA\u00a0 01605 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel E","family":"Bauer","sequence":"additional","affiliation":[{"name":"Division of Hematology\/Oncology, Boston Children\u2019s Hospital , 1 Blackfan Circle,\u00a0Boston, MA 02115 ,","place":["USA"]},{"name":"Department of Pediatric Oncology, Dana-Farber Cancer Institute , 450 Brookline Avenue,\u00a0Boston, MA 02215 ,","place":["USA"]},{"name":"Department of Pediatrics, Harvard Stem Cell Institute, Broad Institute of Harvard and MIT, Harvard Medical School , 25 Shattuck Street, Boston, MA 02115 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nese","family":"Kurt\u00a0Yilmaz","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School , 364 Plantation Street,\u00a0 Worcester , MA\u00a0 01605 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7042-201X","authenticated-orcid":false,"given":"Scot A","family":"Wolfe","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School , 364 Plantation Street,\u00a0 Worcester , MA\u00a0 01605 ,","place":["USA"]},{"name":"Department of Molecular, Cell and Cancer Biology, UMass Chan Medical School , 364 Plantation Street,\u00a0Worcester, MA 01605 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2270-6613","authenticated-orcid":false,"given":"Celia A","family":"Schiffer","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School , 364 Plantation Street,\u00a0 Worcester , MA\u00a0 01605 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,12,16]]},"reference":[{"key":"2025011110012665700_B1","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1038\/nature17946","article-title":"Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage","volume":"533","author":"Komor","year":"2016","journal-title":"Nature"},{"key":"2025011110012665700_B2","doi-asserted-by":"crossref","first-page":"aaf8729","DOI":"10.1126\/science.aaf8729","article-title":"Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems","volume":"353","author":"Nishida","year":"2016","journal-title":"Science"},{"key":"2025011110012665700_B3","doi-asserted-by":"crossref","first-page":"770","DOI":"10.1038\/s41576-018-0059-1","article-title":"Base editing: precision chemistry on the genome and transcriptome of living cells","volume":"19","author":"Rees","year":"2018","journal-title":"Nat. Rev. Genet."},{"key":"2025011110012665700_B4","doi-asserted-by":"crossref","first-page":"1121","DOI":"10.1016\/j.tibtech.2019.03.008","article-title":"CRISPR\/Cas-mediated base editing: technical considerations and practical applications","volume":"37","author":"Molla","year":"2019","journal-title":"Trends Biotechnol."},{"key":"2025011110012665700_B5","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1038\/s41596-020-00450-9","article-title":"Precision genome editing using cytosine and adenine base editors in mammalian cells","volume":"16","author":"Huang","year":"2021","journal-title":"Nat. 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