- 作者: Jun-Xia Deng, Xiao-Jing Nie, Ying-Feng Lei, Chao-Feng Ma, Dong-Liang Xu, Biao Li, Zhi-Kai Xu and Guo-Cheng Zhang
- 作者服務機構: Department of Pediatrics, Xijing Hospital, Fourth Military Medical University, Xi′an, Mainland China
- 中文摘要: --
- 英文摘要:
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[PRIME]-modified (2[PRIME]-O-methylation or 2[PRIME]-fluoro modification) siRNAs were designed to target highly conserved 5[PRIME] 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[PRIME] 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[PRIME]-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[PRIME] UTR that can be targeted to effectively inhibit EV71 replication through RNAi strategies. Appropriate 2[PRIME]-modified siRNAs provide a promising approach to optimizing siRNAs to overcome barriers on RNAi-based antiviral therapies for broader administration. - 中文關鍵字: --
- 英文關鍵字: Enterovirus 71, RNA interference, siRNA, 5′ UTR, Viral inhibition, Antiviral agent, 2′-Omethylation modification, 2′-fluoro modification