Coupling metabolic enhancement to plasmid spread enables programmable antimicrobial control
6 affiliations
- 1Institut Pasteur, Université de Paris, Unité Plasticité du Génome Bactérien, et CNRS, UMR3525 , Paris 75015 ,
- 2Consejo Superior de Investigaciones Científicas (CSIC), Centro Nacional de Biotecnología (CNB), Department of Microbial Biotechnology , Madrid 28049 ,
- 3Chair of Intestinal Microbiome, Technical University of Munich , Freising 85354 ,
- 4Institut Pasteur Université Paris Cité, CNRS UMR 6047, Genetics of Biofilms laboratory , Paris 75015 ,
- 5Collège Doctoral, Sorbonne Université , Paris 75006 ,
- 6German Center for Infection Research (DZIF), Partner site TUM , Munich 80539 ,
Abstract
The rise of multidrug-resistant pathogens underscores the need for precise antimicrobial strategies beyond conventional antibiotics. Conjugation-based approaches offer a powerful yet underexploited means of delivering targeted genetic interventions within microbial communities. Here, we combine selective killing modules with optimized conjugative vectors to target antibiotic-resistant pathogens and clinically relevant resistance plasmids. First, we engineered toxin–intein modules, programmable cassettes that restrict toxic activity to specific regulatory contexts. We developed modules targeting Shigella spp., Salmonella enterica, and bacteria carrying the resistance plasmid pOXA-48, demonstrating tunable and selective activity at both species and strain levels. To identify the optimal delivery platform, we compared mobilizable and conjugative plasmids, finding that conjugative systems consistently outperform mobilizable ones by approximately one order of magnitude. We then streamlined the broad-host-range plasmid RP4 and enhanced its functionality by incorporating the metabolic fos locus, a type IV pilus operon or both features combined. Integration of the anti-pOXA-48 module into these vectors enabled effective suppression of resistance plasmid spread in complex microbial communities. Together, these results advance the use of conjugative plasmids as programmable antimicrobial platforms and identify delivery efficiency and ecological competitiveness as key determinants of the success of such interventions.
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BibTeX
@article{doradomorales2026coupling,
title = {{Coupling metabolic enhancement to plasmid spread enables programmable antimicrobial control}},
author = {Pedro Dorado-Morales and Alicia Calvo-Villamañán and Lisa Niedermeier and Bianca Audrain and Morgan Lambérioux and Magaly Ducos-Galand and Jean-Marc Ghigo and Bärbel Stecher and Álvaro San Millán and Didier Mazel},
journal = {Nucleic Acids Research},
year = {2026},
month = nov,
doi = {10.1093/nar/gkag1100},
url = {https://academic.oup.com/nar/article/doi/10.1093/nar/gkag1100/9011586},
note = {CC BY-NC 4.0},
}
RIS
TY - JOUR
TI - Coupling metabolic enhancement to plasmid spread enables programmable antimicrobial control
AU - Pedro Dorado-Morales
AU - Alicia Calvo-Villamañán
AU - Lisa Niedermeier
AU - Bianca Audrain
AU - Morgan Lambérioux
AU - Magaly Ducos-Galand
AU - Jean-Marc Ghigo
AU - Bärbel Stecher
AU - Álvaro San Millán
AU - Didier Mazel
PY - 2026
DA - 2026/11/04
JO - Nucleic Acids Research
DO - 10.1093/nar/gkag1100
UR - https://academic.oup.com/nar/article/doi/10.1093/nar/gkag1100/9011586
AB - The rise of multidrug-resistant pathogens underscores the need for precise antimicrobial strategies beyond conventional antibiotics. Conjugation-based approaches offer a powerful yet underexploited means of delivering targeted genetic interventions within microbial communities. Here, we combine selective killing modules with optimized conjugative vectors to target antibiotic-resistant pathogens and clinically relevant resistance plasmids. First, we engineered toxin–intein modules, programmable cassettes that restrict toxic activity to specific regulatory contexts. We developed modules targeting Shigella spp., Salmonella enterica, and bacteria carrying the resistance plasmid pOXA-48, demonstrating tunable and selective activity at both species and strain levels. To identify the optimal delivery platform, we compared mobilizable and conjugative plasmids, finding that conjugative systems consistently outperform mobilizable ones by approximately one order of magnitude. We then streamlined the broad-host-range plasmid RP4 and enhanced its functionality by incorporating the metabolic fos locus, a type IV pilus operon or both features combined. Integration of the anti-pOXA-48 module into these vectors enabled effective suppression of resistance plasmid spread in complex microbial communities. Together, these results advance the use of conjugative plasmids as programmable antimicrobial platforms and identify delivery efficiency and ecological competitiveness as key determinants of the success of such interventions.
SN - 0305-1048
SN - 1362-4962
ER -