|
|
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 |
|
|
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.
|
Received: 29 January 2013
Published: 25 May 2013
|
|
|
|
[1] Franklin. D.Lowy. Staphylococcus aureus infections. The New England Journal of Medicine,1998,8(20):520-532. [2] Vipin C K, Purohit H J. Quenching the quorum sensing system: potential antibacterial drug targets. Critical Reviews in Microbiology,2011,37(2): 121-140. [3] 于冰,杨光,邵宁生. 金黄色葡萄球菌中二元调控系统agr及与其他二元系统相互作用的研究进展.军事医学科学院院刊, 2007,31(2):187-190. Yu B, Yang G, Shao N S. Advances in the research on two-component regulatory system, agr, and its interaction with other two-component systems in Staphylococcus aureus. Bulletin of the Academy of Military Medical Sciences, 2007,31(2):187-190. [4] 周颖,薛小燕,白卉,等. 计算机辅助设计抗MRSA耐药基因mecR1的10-23型脱氧核酶.中国药科大学学报,2010,41(4):380-384. Zhou Y, Xue X X, Bai H,et al. Computer-aided design of the 10--23 deoxyribozyme targeting resistance gene mecR1. Journal of China Pharmaceutical University,2010, 41(4):380-384. [5] Rasko D A, Sperandio V. Anti-virulence strategies to combat bacteria-mediated disease. Nat Rev Drug Discov, 2010; 9: 117-128. [6] Raina S, De Vizio D, Odell M, et al. Microbial quorum sensing: a tool or a target for antimicrobial therapy? Biotechnol Appl Biochem, 2009; 54: 65-84. [7] Ng W L, Bassler B L. Bacterial quorum-sensing network architectures. Annu Rev Genet, 2009; 43: 197-222. [8] Park J, Jagasia R, Kaufmann G F,et al. Infection control by antibody disruption of bacterial quorum sensing signaling. Chemistry & Biology,2007, 14(10):1119-1127. [9] Chan W C, Coyle B J, Williams P. Virulence regulation and quorum sensing in staphylococcal infections: competitive agrC antagonists as quorum sensing inhibitors. Journal of Medicinal Chemistry, 2004,47(19):4633-4641. [10] Christensen U, Jacobsen N, Rajwanshi V K, et al. Stopped-flow kinetics of locked nucleic acid (LNA)-oligonucleotide duplex formation: studies of LNA-DNA and DNA-DNA interactions. Biochem J, 2001; 354: 481-4. [11] Good L, Awasthi S K, Dryselius R, et al. Bactericidal antisense effects of peptide-PNA conjugates. 2001,Nat Biotechnol,19: 360-364. [12] Hui Bai, Guojun Sang,Yu You,et al. Targeting RNA polymerase primary σ70 as a therapeutic strategy against methicillin-resistant staphylococcus aureus by Antisense Peptide Nucleic Acid.Plos one,2012,7(1):1-10. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
|
Shared |
|
|
|
|
|
Discussed |
|
|
|
|