Subtractive modification of bacterial consortium using antisense peptide nucleic acids

Area of Science:

• Microbiology
• Synthetic Biology
• Biotechnology

Background:

• Microbiome engineering aims to design and modify microbial communities for specific functions.
• Subtractive microbiome modification requires precise removal of target species.
• Existing methods lack specificity and universal applicability for single-species removal.

Purpose of the Study:

• To evaluate the efficacy of antisense peptide nucleic acid conjugated with cell-penetrating peptides (CPP-PNA) as microbiome modifiers.
• To determine if CPP-PNA can specifically inhibit target bacterial species within a complex consortium.
• To assess the impact of targeted species inhibition on the growth of other community members.

Main Methods:

• Development of a CPP-PNA conjugate designed to target specific bacterial species.
• Application of CPP-PNA to an artificial bacterial consortium containing *Escherichia coli*, *Pseudomonas putida*, *Pseudomonas fluorescens*, and *Lactiplantibacillus plantarum*.
• Monitoring of bacterial growth inhibition and community dynamics following CPP-PNA treatment.

Main Results:

• CPP-PNA specifically inhibited the growth of *Escherichia coli* and *Pseudomonas putida* in the artificial consortium.
• Inhibition of *P. putida* led to increased growth of *P. fluorescens*.
• The removal of *P. putida* also resulted in the inhibition of *L. plantarum* growth.

Conclusions:

• CPP-PNA serves as a potent tool for precise microbiome engineering by enabling targeted species removal.
• This approach facilitates the analysis of inter-species growth relationships and ecological dynamics within microbial communities.
• CPP-PNA holds promise for developing novel strategies in microbiome modulation and synthetic biology applications.