The highly conserved 5' untranslated region as an effective target towards the inhibition of Enterovirus 71 replication by unmodified and appropriate 2'-modified siRNAs

Journal of Biomedical Science

Guo-Cheng Zhang²  Zhi-Kai Xu¹  Biao Li²  Dong-Liang Xu²  Chao-Feng Ma³  Ying-Feng Lei¹  Xiao-Jing Nie²  Jun-Xia Deng² 

[1] Department of Microbiology, Fourth Military Medical University, 17th Changlexi Road, Xi′an, 710032, P. R. China;Department of Pediatrics, Xijing Hospital, Fourth Military Medical University, 15th Changlexi Road, Xi′an, 710032, P. R. China;Department of Viral Diseases Laboratory, Xi′an Center for Disease Control and Prevention, 599th Xiying Road, Xi′an, 710054, P. R. China

关键词: 2^′-fluoro modification;    2^′-O-methylation modification;    Antiviral agent;    Viral inhibition;    5^′ UTR;    siRNA;    RNA interference;    Enterovirus 71;   

Others  :  824666
DOI  :  10.1186/1423-0127-19-73

 received in 2012-04-22, accepted in 2012-08-08,  发布年份 2012

【 摘 要 】

Background

Enterovirus 71 (EV71) is a highly infectious agent that plays an etiological role in hand, foot, and mouth disease. It is associated with severe neurological complications and has caused significant mortalities in recent large-scale outbreaks. Currently, no effective vaccine or specific clinical therapy is available against EV71.

Methods

Unmodified 21 nucleotide small interfering RNAs (siRNAs) and classic 2^′-modified (2^′-O-methylation or 2^′-fluoro modification) siRNAs were designed to target highly conserved 5^′ untranslated region (UTR) of the EV71 genome and employed as anti-EV71 agents. Real-time TaqMan RT-PCR, western blot analysis and plaque assays were carried out to evaluate specific viral inhibition by the siRNAs.

Results

Transfection of rhabdomyosarcoma (RD) cells with siRNAs targeting the EV71 genomic 5^′ UTR significantly delayed and alleviated the cytopathic effects of EV71 infection, increased cell viability in EV71-infected RD cells. The inhibitory effect on EV71 replication was sequence-specific and dosage-dependent, with significant corresponding decreases in viral RNA, VP1 protein and viral titer. Appropriate 2^′-modified siRNAs exhibited similar RNA interference (RNAi) activity with dramatically increased serum stability in comparison with unmodified counterparts.

Conclusion

Sequences were identified within the highly conserved 5^′ UTR that can be targeted to effectively inhibit EV71 replication through RNAi strategies. Appropriate 2^′-modified siRNAs provide a promising approach to optimizing siRNAs to overcome barriers on RNAi-based antiviral therapies for broader administration.

【 授权许可】

   
2012 Deng et al.; licensee BioMed Central Ltd.

【 图 表 】

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