| Literature DB >> 32284602 |
Jan Mathony1,2, Zander Harteveld3,4, Carolin Schmelas5,6, Julius Upmeier Zu Belzen1,2,7, Sabine Aschenbrenner1,2,8, Wei Sun9,10, Mareike D Hoffmann1,8, Christina Stengl1, Andreas Scheck3,4, Sandrine Georgeon3,4, Stéphane Rosset3,4, Yanli Wang9,10, Dirk Grimm5,6,11, Roland Eils2,7, Bruno E Correia12,13, Dominik Niopek14,15.
Abstract
Anti-CRISPR (Acr) proteins are powerful tools to control CRISPR-Cas technologies. However, the available Acr repertoire is limited to naturally occurring variants. Here, we applied structure-based design on AcrIIC1, a broad-spectrum CRISPR-Cas9 inhibitor, to improve its efficacy on different targets. We first show that inserting exogenous protein domains into a selected AcrIIC1 surface site dramatically enhances inhibition of Neisseria meningitidis (Nme)Cas9. Then, applying structure-guided design to the Cas9-binding surface, we converted AcrIIC1 into AcrIIC1X, a potent inhibitor of the Staphylococcus aureus (Sau)Cas9, an orthologue widely applied for in vivo genome editing. Finally, to demonstrate the utility of AcrIIC1X for genome engineering applications, we implemented a hepatocyte-specific SauCas9 ON-switch by placing AcrIIC1X expression under regulation of microRNA-122. Our work introduces designer Acrs as important biotechnological tools and provides an innovative strategy to safeguard CRISPR technologies.Entities:
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Year: 2020 PMID： 32284602 DOI： 10.1038/s41589-020-0518-9
Source DB: PubMed Journal: Nat Chem Biol ISSN： 1552-4450 Impact factor: 15.040