Informace o publikaci
Structural engineering of staphylococcal Kayvirus using CRISPR-Cas10 for imaging and biosensing
| Autoři | |
|---|---|
| Rok publikování | 2025 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | Recent advances in CRISPR-Cas genome editing have expanded the scope of bacteriophage engineering, enabling innovations in medicine, nanotechnology, and synthetic biology. While staphylococcal phage genomes have been modified previously, targeted structural protein engineering has remained largely unexplored. Here, we used a two-strain strategy combining homologous recombination with CRISPR-Cas10-assisted counter-selection to insert a poly-histidine tag into a surface-exposed, flexible loop of the tail sheath protein of Staphylococcus phage 812h1 (Kayvirus). Guided by structure–function relationships, the site was chosen to preserve assembly and infectivity. The His-tag enabled antibody-mediated detection and precise functionalization of the phage surface. Functional validation by bio-layer interferometry, ELISA, flow cytometry, and fluorescence microscopy confirmed that the modification maintained biological activity. The labeled phages were effectively immobilized on biosensor platforms, visualized microscopically, and applied to flow cytometric bacterial identification. These findings underscore the importance of high-resolution structural data for reporter tag design and establish CRISPR-Cas10-mediated structural engineering of Kayvirus phages as a versatile platform for generating application-specific viral particles for diagnostics, biosensing, and potentially therapeutics. |
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