Inhibition of the foot-and-mouth disease virus subgenomic replicon by RNA aptamers.
Inhibition of the foot-and-mouth disease virus subgenomic replicon by RNA aptamers.
复制标题
DOI：
10.1099/vir.0.067751-0
复制
发表时间：
2014-12
期刊：
影响因子：
--
通讯作者：
Stonehouse NJ
中科院分区：
文献类型：
--
作者：
Forrest S;Lear Z;Herod MR;Ryan M;Rowlands DJ;Stonehouse NJ
我们之前已经记录了RNA适体对口蹄疫病毒（3Dpol）的RNA依赖性RNA聚合酶的抑制活性。在这里，我们报告了它们的修饰以及与包含 GFP 的亚基因组复制子（pGFP-PAC 复制子）的使用，从而可以实时监控和量化复制。转染的 BHK-21 细胞中的 GFP 表达在转染后约 8 小时达到最大值，此时细胞形态的变化与病毒诱导的细胞病变效应一致。然而，用 3Dpol 适体 RNA 转染带有复制子的细胞会抑制 GFP 表达并维持正常细胞形态，而对照适体 RNA 几乎没有影响。这种抑制与 3Dpol 的减少（通过免疫印迹检测）相关，并显示出剂量依赖性。 3Dpol 适体似乎比 2'-C-甲基胞苷 (2'CMC) 更有效。因此，复制复合物成分的适体是研究病毒复制的有用分子工具，并且在未来也有潜力作为诊断分子。
We have previously documented the inhibitory activity of RNA aptamers to the RNA-dependent RNA polymerase of foot-and-mouth disease virus (3Dpol). Here we report their modification and use with a subgenomic replicon incorporating GFP (pGFP-PAC replicon), allowing replication to be monitored and quantified in real-time. GFP expression in transfected BHK-21 cells reached a maximum at approximately 8 h post-transfection, at which time change in morphology of the cells was consistent with a virus-induced cytopathic effect. However, transfection of replicon-bearing cells with a 3Dpol aptamer RNA resulted in inhibition of GFP expression and maintenance of normal cell morphology, whereas a control aptamer RNA had little effect. The inhibition was correlated with a reduction in 3Dpol (detected by immunoblotting) and shown to be dose dependent. The 3Dpol aptamers appeared to be more effective than 2′-C-methylcytidine (2′CMC). Aptamers to components of the replication complex are therefore useful molecular tools for studying viral replication and also have potential as diagnostic molecules in the future.
登录
查看更多内容
影响因子：
4.8
作者：
Ferrer-Orta, C;Arias, A;Verdaguer, N
通讯作者：
Verdaguer, N
影响因子：
5.4
作者：
BELSHAM, GJ;BRANGWYN, JK
通讯作者：
BRANGWYN, JK
影响因子：
5.6
作者：
GREEN, L;WAUGH, S;TUERK, C
通讯作者：
TUERK, C
影响因子：
4.8
作者：
Ruckman, J;Green, LS;Janjic, N
通讯作者：
Janjic, N
影响因子：
6.8
作者：
Ferrer-Orta, Cristina;Agudo, Ruben;Verdaguer, Nuria
通讯作者：
Verdaguer, Nuria