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

Area of Science:

• Microbiology
• Genetic Engineering
• Antimicrobial Resistance

Background:

• Bacteriophages offer a promising alternative to traditional antibiotics against resistant bacteria.
• Reprogramming bacteriophages enables targeted delivery of antimicrobial agents.
• Developing novel delivery systems is crucial for combating antimicrobial resistance.

Purpose of the Study:

• To develop a bacteriophage P4 cosmid system for delivering Cas9 antimicrobial agents.
• To engineer bacteriophages for efficient delivery into specific gut pathogens.
• To evaluate the efficacy of the system against Shigella flexneri and Escherichia coli O157:H7.

Main Methods:

• Developed a bacteriophage P4 cosmid system for Cas9 antimicrobial delivery.
• Engineered phage tail fibers to enhance transduction efficiency and host recognition.
• Tested the system's efficacy in delivering Cas9 into S. flexneri and E. coli O157:H7.

Main Results:

• The P4 cosmid system achieved high titers of cosmid-transducing units without helper phage contamination.
• Genetic modification of phage tail fibers improved cosmid DNA transduction in S. flexneri and enabled recognition of E. coli O157:H7.
• Transducing units with chimeric tails significantly enhanced Cas9-mediated killing of both target pathogens.

Conclusions:

• The P4 cas9 cosmid system demonstrates potential as a sequence-specific antimicrobial delivery platform.
• This engineered bacteriophage system shows promise for therapeutic and biocontrol applications against gut pathogens.
• The study highlights the versatility of bacteriophages in combating antimicrobial-resistant bacteria.