Silencing of Pseudomonas aeruginosa Integron Class I Gene by Short Interference RNA

Abstract Background: The elevation of global resistance of Pseudomonas aeruginosa to various antibiotic agents represents a public health threat. In the field of bacterial cellular engineering, gene silencing emerges as an important technology capable of reducing the expression of antibiotic resistance genes. Specifically, short interference RNA (siRNA) technology holds promise in addressing bacterial infections linked to antibiotic resistance. This study focuses on the application of siRNA to target the Integron Class I (intI) gene in P. aeruginosa as a potential strategy to attenuate antibiotic resistance issues. Objectives: The primary objectives of this research involved the identification of the intI gene in P. aeruginosa clinical isolates, the design and introduction of constructed siRNA to induce gene silencing, and the assessment of alterations in gene expression using RT-PCR. The study aims to clarify the efficacy of siRNA-mediated silencing in downregulating the expression of the intI gene associated with antibiotic resistance. Materials and Methods: A total of 30 clinical isolates of P. aeruginosa were included in the study. DNA extraction was performed to identify the intI gene, and a custom siRNA sequence was designed against intI. The siRNA constructs were introduced into P. aeruginosa isolates via Gold Nanoparticles (AuNPs). Gene expression changes were quantified using RT-PCR. Results: The study found that the intI gene was present in all (100%) P. aeruginosa isolates. Importantly, the results demonstrated a significant reduction in the expression level of the intI gene when targeted with siRNA, indicating the efficacy of siRNA in downregulating the gene associated with antibiotic resistance. Conclusion: In conclusion, the widespread presence of the intI gene in P. aeruginosa is closely linked to antibiotic resistance. The study highlights the substantial capability of siRNA to downregulate intI gene expression, presenting a promising avenue to inhibit antibiotic resistance in vitro . This research underscores the potential of siRNA technology as a targeted approach to combat antibiotic resistance in P. aeruginosa .