| 초록 |
Recently, CRISPR/Cas9-mediated genome editing system has enabled the rapid and accurate alterations of genomic information in various organisms. The genomic editing protein Cas9 can be functionalized with various bioactive molecules, making it possible to apply a variety of CRISPR systems for both industrial and medical purposes. Herein, we introduce chemoselective functionalities into the Cas9 protein through biorthogonal noncannonical amino acid tagging. We used azidohomoalanine (AHA) to replace the methionine group of Cas9, which can be further labeled by an azide-alkyne cycloaddition reaction with dibenzocyclooctyne. The high reaction rates and its desired biocompatibility allows Cas9 to be functionalized for therapeutic purposes via nonviral procedures. This approach will have great potential for creating bioorthogonally functionalized Cas9 which can widen the application of CRISPR, in particular for the development of therapeutic proteins with new and improved properties. |
