CRISPR-ASSISTED STRUCTURAL MODIFICATION OF STAPHYLOCOCCAL
BACTERIOPHAGES FOR IMAGING AND BIOSENSING APPLICATIONS

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CRISPR-ASSISTED STRUCTURAL MODIFICATION OF STAPHYLOCOCCAL BACTERIOPHAGES FOR IMAGING AND BIOSENSING ...

Přehled o publikaci

a 2025

CRISPR-ASSISTED STRUCTURAL MODIFICATION OF STAPHYLOCOCCAL BACTERIOPHAGES FOR IMAGING AND BIOSENSING APPLICATIONS

ŠIMEČKOVÁ, Hana; Pavol BÁRDY; Lucie KUNTOVÁ; Eliška MACHÁČOVÁ; Tibor BOTKA et. al.

Basic information

Original name

CRISPR-ASSISTED STRUCTURAL MODIFICATION OF STAPHYLOCOCCAL BACTERIOPHAGES FOR IMAGING AND BIOSENSING APPLICATIONS

Authors

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

Edition

4th Annual Meeting of the National Institute of Virology and Bacteriology (NIVB), Kutná Hora, 2025

Other information

Language

English

Type of outcome

Konferenční abstrakta

Country of publisher

Czech Republic

Confidentiality degree

is not subject to a state or trade secret

References:

URL

Organization

Středoevropský technologický institut – Repository – Repository

ISSN

Keywords in English

CRISPR; Staphylococcus aureus phage; His-phages; microscopy; visualization

Links

LX22NPO5103, research and development project.

Changed: 3/12/2025 00:51, RNDr. Daniel Jakubík

Abstract

In the original language

Lytic bacteriophages are effective therapeutic agents against antibiotic-resistant bacteria. Furthermore, recent advances in CRISPR-Cas–based genome editing techniques have expanded the possibilities of engineering phages with novel and unique properties. In our study, the polyvalent Staphylococcus aureus phage 812h1 was genetically modified by inserting a polyhistidine tag into an exposed loop of the tail sheath protein. An editing strategy combining homologous recombination with CRISPR-Cas10-assisted countercounter-selection allowed the construction of stable recombinant particles. The His-phages were specifically recognized by antibodies, and their attachment to bacteria was visualized by fluorescence microscopy. The structural modifications did not impair the biological activity of the phages, and their functionality was further validated using bio-layer interferometry, enzyme-linked immunosorbent assay, and flow cytometry. The engineered phages provide new opportunities for applications in research, diagnostics, environmental monitoring, and the development of advanced biosensing tools.

Files attached

https://is.muni.cz/publication/2535720/Sbornik_NIVB_meeting_2025.pdf

Cite

ŠIMEČKOVÁ, Hana; Pavol BÁRDY; Lucie KUNTOVÁ; Eliška MACHÁČOVÁ; Tibor BOTKA; Ján BÍŇOVSKÝ; Josef HOUSER; Zdeněk FARKA; Pavel PLEVKA; Roman PANTŮČEK and Ivana MAŠLAŇOVÁ. CRISPR-ASSISTED STRUCTURAL MODIFICATION OF STAPHYLOCOCCAL BACTERIOPHAGES FOR IMAGING AND BIOSENSING APPLICATIONS. In 4th Annual Meeting of the National Institute of Virology and Bacteriology (NIVB), Kutná Hora. 2025. ISSN 2336-7202.

@proceedings{90750,
   author = {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 and Mašlaňová, Ivana},
   booktitle = {4th Annual Meeting of the National Institute of Virology and Bacteriology (NIVB), Kutná Hora},
   keywords = {CRISPR; Staphylococcus aureus phage; His-phages; microscopy; visualization},
   language = {eng},
   title = {CRISPR-ASSISTED STRUCTURAL MODIFICATION OF STAPHYLOCOCCAL BACTERIOPHAGES FOR IMAGING AND BIOSENSING APPLICATIONS},
   url = {https://nivb.cz/en/2025/10/29/nivb-meeting-2025/},
   year = {2025}
}

TY  - CONF
ID  - 90750
AU  - Šimečková, Hana - Bárdy, Pavol - Kuntová, Lucie - Macháčová, Eliška - Botka, Tibor - Bíňovský, Ján - Houser, Josef - Farka, Zdeněk - Plevka, Pavel - Pantůček, Roman - Mašlaňová, Ivana
PY  - 2025
TI  - CRISPR-ASSISTED STRUCTURAL MODIFICATION OF STAPHYLOCOCCAL BACTERIOPHAGES FOR IMAGING AND BIOSENSING APPLICATIONS
KW  - CRISPR
KW  - Staphylococcus aureus phage
KW  - His-phages
KW  - microscopy
KW  - visualization
UR  - https://nivb.cz/en/2025/10/29/nivb-meeting-2025/
N2  - Lytic bacteriophages are effective therapeutic agents against antibiotic-resistant bacteria. Furthermore, recent advances in CRISPR-Cas–based genome editing techniques have expanded the possibilities of engineering phages with novel and unique properties. In our study, the polyvalent Staphylococcus aureus phage 812h1 was genetically modified by inserting a polyhistidine tag into an exposed loop of the tail sheath protein. An editing strategy combining homologous recombination with CRISPR-Cas10-assisted countercounter-selection allowed the construction of stable recombinant particles. The His-phages were specifically recognized by antibodies, and their attachment to bacteria was visualized by fluorescence microscopy. The structural modifications did not impair the biological activity of the phages, and their functionality was further validated using bio-layer interferometry, enzyme-linked immunosorbent assay, and flow cytometry. The engineered phages provide new opportunities for applications in research, diagnostics, environmental monitoring, and the development of advanced biosensing tools.
ER  -

ŠIMEČKOVÁ, Hana; Pavol BÁRDY; Lucie KUNTOVÁ; Eliška MACHÁČOVÁ; Tibor BOTKA; Ján BÍŇOVSKÝ; Josef HOUSER; Zdeněk FARKA; Pavel PLEVKA; Roman PANTŮČEK and Ivana MAŠLAŇOVÁ. CRISPR-ASSISTED STRUCTURAL MODIFICATION OF STAPHYLOCOCCAL BACTERIOPHAGES FOR IMAGING AND BIOSENSING APPLICATIONS. In \textit{4th Annual Meeting of the National Institute of Virology and Bacteriology (NIVB), Kutná Hora}. 2025. ISSN~2336-7202.