{"id":"doi:10.1038/ncomms15790","kind":"article","title":"Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery","authors":["Rees HA","Komor AC","Yeh WH","Caetano-Lopes J","Warman M","Edge ASB","Liu DR"],"author_count":7,"year":2017,"published":"2017-06-06","venue":"Nature communications","cited_by":355,"doi":"10.1038/ncomms15790","arxiv_id":null,"nct_id":null,"source_url":"https://doi.org/10.1038/ncomms15790","archive_url":"https://space-kitty.com/papers/doi:10.1038/ncomms15790","updated":null,"pmid":"28585549","pmcid":"PMC5467206","openalex_id":null,"pdf_url":"https://europepmc.org/articles/PMC5467206/pdf","abstract":"We recently developed base editing, a genome-editing approach that enables the programmable conversion of one base pair into another without double-stranded DNA cleavage, excess stochastic insertions and deletions, or dependence on homology-directed repair. The application of base editing is limited by off-target activity and reliance on intracellular DNA delivery. Here we describe two advances that address these limitations. First, we greatly reduce off-target base editing by installing mutations into our third-generation base editor (BE3) to generate a high-fidelity base editor (HF-BE3). Next, we purify and deliver BE3 and HF-BE3 as ribonucleoprotein (RNP) complexes into mammalian cells, establishing DNA-free base editing. RNP delivery of BE3 confers higher specificity even than plasmid transfection of HF-BE3, while maintaining comparable on-target editing levels. Finally, we apply these advances to deliver BE3 RNPs into both zebrafish embryos and the inner ear of live mice to achieve specific, DNA-free base editing in vivo.","abstract_license":"CC-BY (article license as recorded by Europe PMC; abstract reproduced unmodified apart from markup removal)","license":"CC-BY","is_open_access":true,"retracted":false,"peer_reviewed":null,"categories":[],"details":null,"provenance":"europepmc","attribution":"Abstract of \"Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery\" by Rees HA, Komor AC, Yeh WH et al., https://doi.org/10.1038/ncomms15790, used under its Creative Commons Attribution license (CC-BY); markup removed, otherwise unmodified.","topics":[{"slug":"gene-editing-therapies","name":"CRISPR & Gene-Editing Therapies","domain_slug":"medicine"}],"cited_in_dossiers":[]}