AI Article Synopsis
- Researchers modified a protein called AcrIIC1, which inhibits CRISPR-Cas9, to enhance its effectiveness against various types of Cas9.
- By inserting additional protein domains into AcrIIC1, they significantly improved its ability to inhibit Neisseria meningitidis Cas9.
- The new variant, AcrIIC1X, was developed to specifically target Staphylococcus aureus Cas9, with applications in precise genome editing, including a method to control gene expression based on liver-specific microRNA.
Article 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.
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| http://dx.doi.org/10.1038/s41589-020-0518-9 | DOI Listing |
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