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siRNA-mediated gene silencing of MexB from the MexA-MexB-OprM efflux pump in Pseudomonas aeruginosa

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To gain insights into the effect of MexB gene under the shortinterfering RNA (siRNA), we synthesized 21 bp siRNAduplexes against the MexB gene. RT-PCR was performed todetermine whether the siRNA inhibited the expression ofMexB mRNA. Changes in antibiotic susceptibility in responseto siRNA were measured by the E-test method. The efficacy ofsiRNAs was determined in a murine model of chronic P. aeruginosa lung infection. MexB-siRNAs inhibited both mRNAexpression and the activity of P. aeruginosa in vitro. In vivo,siRNA was effective in reducing the bacterial load in themodel of chronic lung infection and the P. aeruginosa-inducedpathological changes. MexB-siRNA treatment enhanced theproduction of inflammatory cytokines in the early infectionstage (P < 0.05). Our results suggest that targeting of MexBwith siRNA appears to be a novel strategy for treating P. aeruginosa infections.

Abstract

To gain insights into the effect of MexB gene under the shortinterfering RNA (siRNA), we synthesized 21 bp siRNAduplexes against the MexB gene. RT-PCR was performed todetermine whether the siRNA inhibited the expression ofMexB mRNA. Changes in antibiotic susceptibility in responseto siRNA were measured by the E-test method. The efficacy ofsiRNAs was determined in a murine model of chronic P. aeruginosa lung infection. MexB-siRNAs inhibited both mRNAexpression and the activity of P. aeruginosa in vitro. In vivo,siRNA was effective in reducing the bacterial load in themodel of chronic lung infection and the P. aeruginosa-inducedpathological changes. MexB-siRNA treatment enhanced theproduction of inflammatory cytokines in the early infectionstage (P < 0.05). Our results suggest that targeting of MexBwith siRNA appears to be a novel strategy for treating P. aeruginosa infections.

참고문헌 (30)

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