Reprogramming Targeted-Antibacterial-Plasmids (TAPs) to achieve broad-host range antibacterial activity

Area of Science:

• Microbiology
• Molecular Biology
• Genetics

Background:

• Antimicrobial resistance (AMR) is a growing global health crisis, rendering antibiotics ineffective against multidrug-resistant bacteria.
• Novel therapeutic strategies are urgently needed to combat bacterial infections and overcome antibiotic resistance.

Purpose of the Study:

• To investigate the potential of mobilizable Targeted-Antibacterial-Plasmids (TAPs) as a strain-specific antimicrobial treatment.
• To expand the host-range of TAPs beyond Enterobacteriaceae to include phylogenetically distant bacterial species.

Main Methods:

• Development of TAPs carrying CRISPR/Cas systems for targeted bacterial killing.
• Utilizing F plasmid conjugation for initial TAP transfer and host targeting.
• Employing the RP4 plasmid conjugation system to broaden TAP host-range and mobilization capabilities.

Main Results:

• TAPs successfully demonstrated strain-specific antibacterial activity against various Enterobacteriaceae, including E. coli K12, EPEC, Enterobacter cloacae, and Citrobacter rodentium.
• Expansion of TAP host-range using RP4 conjugation enabled targeting of diverse species such as Salmonella enterica Typhimurium, Klebsiella pneumoniae, Vibrio cholerae, and Pseudomonas aeruginosa.

Conclusions:

• The TAP strategy is versatile and adaptable for targeting a broad spectrum of bacterial pathogens.
• This research represents a significant advancement toward developing effective, non-antibiotic, strain-specific antimicrobial therapies to combat AMR.