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Sequence-Specific Gene Silencing of acrA in the Multi-drug Efflux System AcrAB Induces Sensitivity in Drug-Resistant Klebsiella pneumoniae

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Abstract

Multi-drug efflux is one of the resistant determinants in Klebsiella pneumoniae that are encountered in a broad range of clinically relevant antimicrobial agents. An alternative method to strategically induce sensitivity in drug-resistant K. pneumoniae and improve the efficacy of the existing antibiotics is the need of the hour. Hence, an antisense RNA was designed against the acrA gene of the AcrAB-TolC efflux system in a drug-resistant isolate of K. pneumoniae obtained from a blood culture. Minimum inhibitory concentration by E test demonstrated that the antisense RNA could significantly increase the susceptibility of previously resistant K. pneumoniae toward ciprofloxacin (CIP) and co-trimoxazole. Real-time PCR determined the ability of the antisense RNA to inhibit the expression of the acrA-mRNA. The wild-type K. pneumoniae showed increased growth in the presence of CIP, while, under the same condition, the growth of the antisense RNA-treated K. pneumoniae was inhibited up till 12 h. In the presence of co-trimoxazole, delayed growth rate of the antisense RNA-treated K. pneumoniae was seen, in comparison to that of the wild-type K. pneumoniae and also a fourfold reduction was noted in the expression of the efflux gene acrA. Our results underscore the potential of the acrA antisense RNA as an alternative therapeutic against multi-drug-resistant K. pneumoniae.

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Acknowledgements

The corresponding author gratefully acknowledges Nitte (Deemed to be University) for the financial support received through research Grant No. NUFR1/2018/10/17, toward this study.

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Authors and Affiliations

  1. Nitte University Centre for Science Education and Research, Nitte (Deemed to be University), Deralakatte, Mangaluru, 575018, India

    Priyanka Ashwath, Vijaya Kumar Deekshit, Anusha Rohit, Praveen Rai, Vankadari Aditya, Nishith Babu, Indrani Karunasagar & Akhila Dharnappa Sannejal

  2. Department of Microbiology, Madras Medical Mission, Chennai, India

    Anusha Rohit

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  1. Priyanka Ashwath
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Contributions

PA contributed to formal analysis and writing of the original draft, DVK contributed to formal analysis and investigation, AR provided isolate source and performed draft reviewing and editing, PR contributed to formal analysis and investigation, VA contributed to formal analysis, NB contributed to formal analysis, IK contributed to final reviewing and editing of the manuscript, ADS contributed to supervision and funding acquisition.

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Correspondence to Akhila Dharnappa Sannejal.

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This study was approved by the Institutional Ethics Committee of the Madras Medical Mission, Chennai (Reg. No.: ECR/140/Inst/TN/2013/RR-20).

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Ashwath, P., Deekshit, V.K., Rohit, A. et al. Sequence-Specific Gene Silencing of acrA in the Multi-drug Efflux System AcrAB Induces Sensitivity in Drug-Resistant Klebsiella pneumoniae. Mol Biotechnol 65, 953–960 (2023). https://doi.org/10.1007/s12033-022-00585-y

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