{"id":"doi:10.1038/s41467-021-22009-2","kind":"article","title":"CRISPR-Cas9 cytidine and adenosine base editing of splice-sites mediates highly-efficient disruption of proteins in primary and immortalized cells","authors":["Kluesner MG","Lahr WS","Lonetree CL","Smeester BA","Qiu X","Slipek NJ","Claudio Vázquez PN","Pitzen SP","Pomeroy EJ","Vignes MJ","Lee SC","Bingea SP","Andrew AA","Webber BR","Moriarity BS"],"author_count":15,"year":2021,"published":"2021-04-23","venue":"Nature communications","cited_by":107,"doi":"10.1038/s41467-021-22009-2","arxiv_id":null,"nct_id":null,"source_url":"https://doi.org/10.1038/s41467-021-22009-2","archive_url":"https://space-kitty.com/papers/doi:10.1038/s41467-021-22009-2","updated":null,"pmid":"33893286","pmcid":"PMC8065034","openalex_id":null,"pdf_url":"https://europepmc.org/articles/PMC8065034/pdf","abstract":"CRISPR-Cas9 cytidine and adenosine base editors (CBEs and ABEs) can disrupt genes without introducing double-stranded breaks by inactivating splice sites (BE-splice) or by introducing premature stop (pmSTOP) codons. However, no in-depth comparison of these methods or a modular tool for designing BE-splice sgRNAs exists. To address these needs, we develop SpliceR ( http://z.umn.edu/spliceR ) to design and rank BE-splice sgRNAs for any Ensembl annotated genome, and compared disruption approaches in T cells using a screen against the TCR-CD3 MHC Class I immune synapse. Among the targeted genes, we find that targeting splice-donors is the most reliable disruption method, followed by targeting splice-acceptors, and introducing pmSTOPs. Further, the CBE BE4 is more effective for disruption than the ABE ABE7.10, however this disparity is eliminated by employing ABE8e. Collectively, we demonstrate a robust method for gene disruption, accompanied by a modular design tool that is of use to basic and translational researchers alike.","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 \"CRISPR-Cas9 cytidine and adenosine base editing of splice-sites mediates highly-efficient disruption of proteins in primary and immortalized cells\" by Kluesner MG, Lahr WS, Lonetree CL et al., https://doi.org/10.1038/s41467-021-22009-2, 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":[]}