The Role of O-antigen in P1 Transduction of Shigella flexneri and Escherichia coli with its Alternative S' Tail Fibre

Huan, Yang Wei; Fa-arun, Jidapha; Wang, Qianqian

doi:10.1016/j.jmb.2022.167829

Cited by 6 publications

(13 citation statements)

References 58 publications

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“…flexneri serotype 2a and 5a share a common O -polysaccharide backbone but with different glucosyl and O -acetyl groups , (Figure b). The difference in the cell surface component such as O-antigen and core lipopolysaccharides (LPS) can influence phage infectivity and host range. − Therefore, we hypothesized that the difference in the O-antigen modification between the two S. flexneri strains could potentially affect transduction of cas9 cosmids, which would contribute to the difference in the Cas9-mediated killing.…”

Section: Resultsmentioning

confidence: 99%

“…One such candidate is bacteriophage P1, which was shown to transduce phagemid DNA into S. flexneri strains at a high efficiency. , P1 expresses two different tail fibers, namely, the S and S′, which allow host tropism switching. − Our previous study demonstrates that the P1 with S′ tail fiber could transduce cas9 constructs into S. flexneri serotypes 2a and 5a at a high efficiency .…”

Section: Resultsmentioning

confidence: 99%

“…flexneri strains at a high efficiency. , P1 expresses two different tail fibers, namely, the S and S′, which allow host tropism switching. − Our previous study demonstrates that the P1 with S′ tail fiber could transduce cas9 constructs into S. flexneri serotypes 2a and 5a at a high efficiency . Therefore, the S′ tail fiber was a promising candidate for constructing a chimeric P2 tail fiber.…”

Section: Resultsmentioning

confidence: 99%

“…The difference in the cell surface component such as O-antigen and core lipopolysaccharides (LPS) can influence phage infectivity and host range. 49 − 53 Therefore, we hypothesized that the difference in the O-antigen modification between the two S. flexneri strains could potentially affect transduction of cas9 cosmids, which would contribute to the difference in the Cas9-mediated killing. Tail fiber/tail spike engineering can expand or alter the host range of the bacteriophage.…”

Section: Resultsmentioning

confidence: 99%

“… 60 − 62 Our previous study demonstrates that the P1 with S′ tail fiber could transduce cas9 constructs into S. flexneri serotypes 2a and 5a at a high efficiency. 49 Therefore, the S′ tail fiber was a promising candidate for constructing a chimeric P2 tail fiber.…”

Section: Resultsmentioning

confidence: 99%

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Tail-Engineered Phage P2 Enables Delivery of Antimicrobials into Multiple Gut Pathogens

2023

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Bacteriophages can be reprogrammed to deliver antimicrobials for therapeutic and biocontrol purposes and are a promising alternative treatment to antimicrobial-resistant bacteria. Here, we developed a bacteriophage P4 cosmid system for the delivery of a Cas9 antimicrobial into clinically relevant human gut pathogens Shigella flexneri and Escherichia coli O157:H7. Our P4 cosmid design produces a high titer of cosmid-transducing units without contamination by a helper phage. Further, we demonstrate that genetic engineering of the phage tail fiber improves the transduction efficiency of cosmid DNA in S. flexneri M90T as well as allows recognition of a nonnative host, E. coli O157:H7. We show that the transducing units with the chimeric tails enhanced the overall Cas9-mediated killing of both pathogens. This study demonstrates the potential of our P4 cas9 cosmid system as a DNA sequence-specific antimicrobial against clinically relevant gut pathogenic bacteria.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Tail-Engineered Phage P2 Enables Delivery of Antimicrobials into Multiple Gut Pathogens

2023

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P1 Bacteriophage-Enabled Delivery of CRISPR-Cas9 Antimicrobial Activity Against Shigella flexneri

et al. 2023

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The discovery of clustered, regularly interspaced, short palindromic repeats (CRISPR) and the Cas9 RNA-guided nuclease provides unprecedented opportunities to selectively kill specific populations or species of bacteria. However, the use of CRISPR-Cas9 to clear bacterial infections in vivo is hampered by the inefficient delivery of cas9 genetic constructs into bacterial cells. Here, we use a broad-host-range P1-derived phagemid to deliver the CRISPR-Cas9 chromosomal-targeting system into Escherichia coli and the dysentery-causing Shigella flexneri to achieve DNA sequence-specific killing of targeted bacterial cells. We show that genetic modification of the helper P1 phage DNA packaging site (pac) significantly enhances the purity of packaged phagemid and improves the Cas9-mediated killing of S. flexneri cells. We further demonstrate that P1 phage particles can deliver chromosomaltargeting cas9 phagemids into S. flexneri in vivo using a zebrafish larvae infection model, where they significantly reduce the bacterial load and promote host survival. Our study highlights the potential of combining P1 bacteriophage-based delivery with the CRISPR chromosomal-targeting system to achieve DNA sequence-specific cell lethality and efficient clearance of bacterial infection.

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Bacterial synthetic biology: tools for novel drug discovery

Wang

Zhou

2023

Expert Opinion on Drug Discovery

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The Role of O-antigen in P1 Transduction of Shigella flexneri and Escherichia coli with its Alternative S' Tail Fibre

Cited by 6 publications

References 58 publications

Tail-Engineered Phage P2 Enables Delivery of Antimicrobials into Multiple Gut Pathogens

Tail-Engineered Phage P2 Enables Delivery of Antimicrobials into Multiple Gut Pathogens

P1 Bacteriophage-Enabled Delivery of CRISPR-Cas9 Antimicrobial Activity Against Shigella flexneri

Bacterial synthetic biology: tools for novel drug discovery

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