Structural engineering of staphylococcal Kayvirus using
CRISPR-Cas10 for imaging and biosensing

a 2025

Structural engineering of staphylococcal Kayvirus using CRISPR-Cas10 for imaging and biosensing

MAŠLAŇOVÁ, Ivana; Hana ŠIMEČKOVÁ; Pavol BÁRDY; Lucie KUNTOVÁ; Eliška MACHÁČOVÁ et al.

Základní údaje

Originální název

Structural engineering of staphylococcal Kayvirus using CRISPR-Cas10 for imaging and biosensing

Autoři

MAŠLAŇOVÁ, Ivana; Hana ŠIMEČKOVÁ; Pavol BÁRDY; Lucie KUNTOVÁ; Eliška MACHÁČOVÁ; Tibor BOTKA; Ján BÍŇOVSKÝ; Josef HOUSER; Zdeněk FARKA; Pavel PLEVKA a Roman PANTŮČEK

Vydání

The Phage Protein Meeting, 2025

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Belgie

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Označené pro přenos do RIV

Ano

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova česky

CRISPR-Cas10 editace; bakteriofág; protilátka

Klíčová slova anglicky

CRISPR-Cas10 editing; bacteriophage; antibody

Změněno: 19. 1. 2026 17:27, Mgr. Ivana Mašlaňová, Ph.D.

Anotace

V originále

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.

Návaznosti

LX22NPO5103, projekt VaV

Název: Národní institut virologie a bakteriologie (Akronym: NIVB)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Národní institut virologie a bakteriologie, 5.1 EXCELES

NU22-05-00042, projekt VaV

Název: Vývoj nových nanobiotechnologií pro sledování terapeutických bakteriofágů v klinických vzorcích

Investor: Ministerstvo zdravotnictví ČR, Vývoj nových nanobiotechnologií pro sledování terapeutických bakteriofágů v klinických vzorcích, Podprogram 1 - standardní

Citovat

MAŠLAŇOVÁ, Ivana; Hana ŠIMEČKOVÁ; Pavol BÁRDY; Lucie KUNTOVÁ; Eliška MACHÁČOVÁ; Tibor BOTKA; Ján BÍŇOVSKÝ; Josef HOUSER; Zdeněk FARKA; Pavel PLEVKA a Roman PANTŮČEK. Structural engineering of staphylococcal Kayvirus using CRISPR-Cas10 for imaging and biosensing. In The Phage Protein Meeting. 2025.

@proceedings{2547098,
   author = {Mašlaňová, Ivana and Šimečková, Hana and Bárdy, Pavol and Kuntová, Lucie and Macháčová, Eliška and Botka, Tibor and Bíňovský, Ján and Houser, Josef and Farka, Zdeněk and Plevka, Pavel and Pantůček, Roman},
   booktitle = {The Phage Protein Meeting},
   keywords = {CRISPR-Cas10 editing; bacteriophage; antibody},
   language = {eng},
   title = {Structural engineering of staphylococcal Kayvirus using CRISPR-Cas10 for imaging and biosensing},
   url = {https://phageproteinmeeting.ugent.be/static/TPPM_abstract_book.pdf},
   year = {2025}
}

TY  - CONF
ID  - 2547098
AU  - Mašlaňová, Ivana - Šimečková, Hana - Bárdy, Pavol - Kuntová, Lucie - Macháčová, Eliška - Botka, Tibor - Bíňovský, Ján - Houser, Josef - Farka, Zdeněk - Plevka, Pavel - Pantůček, Roman
PY  - 2025
TI  - Structural engineering of staphylococcal Kayvirus using CRISPR-Cas10 for imaging and biosensing
KW  - CRISPR-Cas10 editing
KW  - bacteriophage
KW  - antibody
UR  - https://phageproteinmeeting.ugent.be/static/TPPM_abstract_book.pdf
N2  - 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.
ER  -

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