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| Blocking Methicilli-Resistant Staphylococcus Aureus Agr Quorom Sensing System by Antisense LNA |
| DA Fei, HOU Zheng, MENG Jing-ru, JIA Min, LUO Xiao-xing |
| Department of Pharmacology, Fouth Military Medical University, Xi’an 710032, China |
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Abstract Objective: Using antisense strategy to design and synthesize oligonucleotide targeting agrA mRNA to block agr quorum sensing system,so as to depress the virulence gene of MRSA. Methods: We design and select effective antisense locked nucleic acid (LNA) targeting agrA mRNA by software Primer Premier 5.0 and RNA structure 4.5 and covalently conjugated LNA with the cell penetrating peptide (KFF)3K. The transcription of agrA and downstream virulence genes were analyzed by real-time quantitative PCR and the expression of α-toxin was detected by western blot. Results: Neither PLNA34 nor PLNA522 inhibit the growth of MRSA in vitro. Both of PLNA34 and PLNA522 can significantly reduce the transcription of agrA, the effector RNAⅢ and hla, with PLNA34 having more significant inhibitory effect. PLNA34 has a stronger inhibition in expression of α-toxin than PLNA522. Conclusion: agrA could be a promising drug target to combat MRSA infection.
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Received: 29 January 2013
Published: 25 May 2013
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