Sirna-Mediated Gene Silencing of Tetk Drug Efflux Pump in Staphylococcus Aureus as a Revolutionary Strategy to Combat Bacterial Tetracycline Resistance

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Saif Ismael Hussein, Fawzyia A. R Ibrahim, Anas Y. Al-Hayawi

Abstract

       Staphylococcus aureus (S. aureus) is one of the most common bacteria that cause significant illness owing to the development of antibiotic resistance. Antibiotic-resistant microorganisms, such as S. aureus, need novel research and development approaches.  Small interfering RNA (siRNA) duplexes of 21-23 bp were synthesized against the mRNA of the tetK gene in S. aureus to evaluate the effect of siRNA in the tetK gene expression. Reverse transcription PCR (RT-PCR) was used for detection the effect of the siRNA in tetK mRNA expression. The disc diffusion technique was used to assess differences of the tetracycline susceptibility in response to the action of siRNA. In vitro, tetK-siRNAs reduced both mRNA expression and activity of S. aureus. The effectiveness of siRNA was assessed by comparing the antibacterial activity of S. aureus before and after the introduction of tetK-siRNA to the bacterium. Both modified tetK-siRNA sequence and unmodified tetK-siRNA sequence, were used in study. RT-qPCR revealed that two tetK-siRNA sequences strongly suppressed the production of tetK-mRNA, P = (0.04 and 0.03) respectively at (P < 0.05) when in comparison with control. Regarding the effect of treatment with both tetK-siRNA (Modified) and tetK-siRNA (Unmodified) were the same, the S. aureus bacteria with siRNA, there were 2/4 (50%) were sensitive to tetracycline, while 1/4 isolates (25 %) was intermediate to tetracycline, and 1/4 (25 %) were resistant to tetracycline. According to the findings, siRNA targeting of tetK gene looks to be a novel strategy for treating S. aureus illnesses.

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Saif Ismael Hussein, Fawzyia A. R Ibrahim, Anas Y. Al-Hayawi. (2022). Sirna-Mediated Gene Silencing of Tetk Drug Efflux Pump in Staphylococcus Aureus as a Revolutionary Strategy to Combat Bacterial Tetracycline Resistance. Annals of the Romanian Society for Cell Biology, 26(01), 1670–1681. Retrieved from https://annalsofrscb.ro/index.php/journal/article/view/11096
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