| Резюме: |
The dissertation focuses on the possibility of application of antisense oligonucleotides (ASOs) as antibacterial agents in the fight against resistant pathogenic bacteria. The aim of the study was to design chimeric ASOs active against bacterial mRNA encoding the enzyme adenylate kinase (ADK) with antibacterial activity against S. aureus and ASOs active against the thiamine pyrophosphate riboswitch (TPP RS) with antibacterial activity against L. monocytogenes by the means of bioinformatics methods and synchronization of bacterial cultures described. Following bioinformatic analysis, target nucleotide sequences were selected that are not targeted by existing antibiotics, are not found in humans, and are being used for the first time. The efficacy of the designed ACOs was demonstrated in in vitro experiments, which showed that: 1) ACO1 at a concentration of 1000 nM stopped cell division and exerted a bactericidal effect due to specific binding to the target sequence in S. aureus; 2) ASO2 - a unique sequence that is not found in any organism, does not bind to the genome of the treated bacterial cells and has no antibacterial effect; 3) ASO4 inhibits the bacterial growth of L. monocytogenes and B. subtilis, which contain specific target sequences but not E. coli due to the lack of specific binding. The results achieved may contribute to the development of a new antibiotic class in the fight against resistant pathogens.
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