ArticleMethods in molecular biology (Clifton, N.J.)2026
In Vitro Preclinical Evaluation of BPP-PNA Antisense Antibiotics: A Protocol for Efficacy, Safety, and Resistance Analysis.
Article in Methods in molecular biology (Clifton, N.J.), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
The global challenge of antibacterial resistance necessitates urgent efforts to develop novel antimicrobial agents with innovative chemical scaffolds. Bacterial-penetrating peptide-peptide nucleic acid (BPP-PNA) conjugates represent a promising class of antisense antibiotics (ASObiotics) capable of selectively targeting bacterial gene expression. This chapter provides a structured protocol for the preclinical evaluation of new BPP-PNA compounds, focusing on efficacy, resistance potential, safety, and mechanism of action. Efficacy assessment includes determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), followed by time-kill assays to evaluate bactericidal kinetics. The combination efficacy of multiple BPP-PNAs is investigated using checkerboard assays. Safety and biocompatibility studies include hemolysis assays on red blood cells and cytotoxicity assessments using cell lines relevant to the infection site and drug metabolism and clearance. To evaluate the propensity for resistance development (spontaneous mutation frequency), bacterial populations (10
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