Antisense mqsR-PNA as a putative target to the eradication of Pseudomonas aeruginosa persisters

Abstract

Chemotherapy is frequently unsuccessful in fully eradicating bacterial biofilm infections. Persisters are a main cause for the failure of antibiotic therapies and are assumed to significantly impact the increased multidrug tolerance and unsuccessful elimination of chronic biofilm infections. Pseudomonas aeruginosa infections are frequently linked to high rates of drug-tolerant persisters, triggering a major challenge to human health. It is crucial to classify persisters to develop novel useful therapeutic strategies to fight infectious diseases. In this study, the mqsR gene was selected as a novel antimicrobial target, and silencing was with antisense peptide nucleic acid (PNA) assay to eradicate the P. aeruginosa persisters. First, they were analysed by experimental procedures. Functionality was assessed by stress conditions. We found that the expression of mqsR (as the toxin) compared with mqsA (as antitoxin) was increased under stress conditions. We demonstrated that when mqsR was targeted and treated with different concentrations of mqsR-PNA after 24 hours; the formation of P. aeruginosa persisters was eradicated. Antisense mqsR-PNA in concentrations of 35 μM or more could eradicate persister cell formation in P. aeruginosa. It was suggested that other toxin–antitoxin loci in P. aeruginosa are examined by antisense PNA to detect their functionality. However, considering the importance of persisters in human infections, ex vivo, in vivo, preclinical and clinical settings should be highlighted.