4.7 Article

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

期刊

ACS SYNTHETIC BIOLOGY
卷 12, 期 2, 页码 596-607

出版社

AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.2c00615

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bacteriophage P2/P4; phage-based delivery vector; Cas9 antimicrobial; tail fiber engineering; Escherichia coli O157:H7; Shigella flexneri

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This study developed a bacteriophage P4 cosmid system for delivering Cas9 antimicrobial into clinically relevant human gut pathogens. The genetic engineering of the phage tail fiber improved the transduction efficiency of cosmid DNA and allowed recognition of a nonnative host. This research demonstrates the potential of the P4 cas9 cosmid system as a DNA sequence-specific antimicrobial against clinically relevant gut pathogenic bacteria.
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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