Modification of T2 phage infectivity toward<i>Escherichia coli</i>O157:H7 via using CRISPR/Cas9

Hoshiga, Fumiya; Yoshizaki, K.; Takao, Nobumasa; Miyanaga, Kazuhiko; Tanji, Yasunori

doi:10.1093/femsle/fnz041

Cited by 22 publications

(13 citation statements)

References 31 publications

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“…This idea has been developed previously, and various attempts of the use of phage therapy in the potential treatment of infections caused by STEC have been reported. They included the use of various STEC-specific phages used alone on in cocktails, as well as in combination with probiotics [ 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 ]. Moreover, the use of bacteriophages (without combination with other agents) against a biofilm-forming STEC strain has been reported recently [ 40 ].…”

Section: Discussionmentioning

confidence: 99%

Synergistic Effects of Bacteriophage vB_Eco4-M7 and Selected Antibiotics on the Biofilm Formed by Shiga Toxin-Producing Escherichia coli

et al. 2022

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Apart from antibiotic resistance of pathogenic bacteria, the formation of biofilms is a feature that makes bacterial infections especially difficulty to treat. Shiga toxin-producing Escherichia coli (STEC) strains are dangerous pathogens, causing severe infections in humans, and capable of biofilm production. We have reported previously the identification and characterization of the vB_Eco4-M7 bacteriophage, infecting various STEC strains. It was suggested that this phage might be potentially used in phage therapy against these bacteria. Here, we tested the effects of vB_Eco4-M7 alone or in a phage cocktail with another STEC-infecting phage, and/or in a combination with different antibiotics (ciprofloxacin and rifampicin) on biofilm formed by a model STEC strain, named E. coli O157:H7 (ST2-8624). The vB_Eco4-M7 phage appeared effective in anti-biofilm action in all these experimental conditions (2–3-fold reduction of the biofilm density, and 2–3 orders of magnitude reduction of the number of bacterial cells). However, the highest efficiency in reducing a biofilm’s density and number of bacterial cells was observed when phage infection preceded antibiotic treatment (6-fold reduction of the biofilm density, and 5–6 orders of magnitude reduction of the number of bacterial cells). Previous reports indicated that the use of antibiotics to treat STEC-caused infections might be dangerous due to the induction of Shiga toxin-converting prophages from bacterial genomes under stress conditions caused by antibacterial agents. We found that ciprofloxacin was almost as efficient in inducing prophages from the E. coli O15:H7 (ST2-8624) genome as a classical inducer, mitomycin C, while no detectable prophage induction could be observed in rifampicin-treated STEC cells. Therefore, we conclude the latter antibiotic or similarly acting compounds might be candidate(s) as effective and safe drug(s) when used in combination with phage therapy to combat STEC-mediated infections.

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Section: Discussionmentioning

confidence: 99%

Synergistic Effects of Bacteriophage vB_Eco4-M7 and Selected Antibiotics on the Biofilm Formed by Shiga Toxin-Producing Escherichia coli

et al. 2022

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“…The CRISPR-Cas system has been widely used to not only target desired mutant phages but also to modify the phages in vivo, with the Type I CRISPR system used to modify phages of bacteriophage T7 [14] and lytic phage ICP1 [36]. Phages altered with the Type II system include phage P2 of Lactococcus lactis, phiKpS2 of Klebsiella pneumoniae, and phages T7, T4, T2, and KF1 of Escherichia coli [37][38][39][40]. Type III systems have been used to alter phages of Staphylococcus epidermidis and Staphylococcus aureus [41].…”

Section: Advancesmentioning

confidence: 99%

Conceptual Framework and Advances in CRISPR-Cas Based and BRED Phage Engineering

Sharma¹

2021

IJSRBS

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“…A different type I CRISPR system has also been used to create mutants of the lytic phage ICP1 that infects V. cholerae [26]. A number of phages have been modified with type II CRISPR systems; including phage 2972 infecting Streptococcus thermophilus, phage P2 infecting Lactococcus lactis, phiKpS2 infecting Klebsiella pneumoniae and phages T2, T4, T7 and KF1 that infect E. coli respectively [25,[27][28][29][30][31]. A type III CRISPR system has also been used to engineer phages that infect Staphylococcus aureus and Staphylococcus epidermidis [32].…”

Section: Introductionmentioning

confidence: 99%

Comparison of CRISPR and Marker-Based Methods for the Engineering of Phage T7

Grigonyte

Harrison

MacDonald

et al. 2020

Viruses

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With the recent rise in interest in using lytic bacteriophages as therapeutic agents, there is an urgent requirement to understand their fundamental biology to enable the engineering of their genomes. Current methods of phage engineering rely on homologous recombination, followed by a system of selection to identify recombinant phages. For bacteriophage T7, the host genes cmk or trxA have been used as a selection mechanism along with both type I and II CRISPR systems to select against wild-type phage and enrich for the desired mutant. Here, we systematically compare all three systems; we show that the use of marker-based selection is the most efficient method and we use this to generate multiple T7 tail fibre mutants. Furthermore, we found the type II CRISPR-Cas system is easier to use and generally more efficient than a type I system in the engineering of phage T7. These results provide a foundation for the future, more efficient engineering of bacteriophage T7.

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Modification of T2 phage infectivity towardEscherichia coliO157:H7 via using CRISPR/Cas9

Cited by 22 publications

References 31 publications

Synergistic Effects of Bacteriophage vB_Eco4-M7 and Selected Antibiotics on the Biofilm Formed by Shiga Toxin-Producing Escherichia coli

Synergistic Effects of Bacteriophage vB_Eco4-M7 and Selected Antibiotics on the Biofilm Formed by Shiga Toxin-Producing Escherichia coli

Conceptual Framework and Advances in CRISPR-Cas Based and BRED Phage Engineering

Comparison of CRISPR and Marker-Based Methods for the Engineering of Phage T7

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