Significant inhibition of Tembusu virus envelope and NS5 gene using an adenovirus-mediated short hairpin RNA delivery system
Significant inhibition of Tembusu virus envelope and NS5 gene using an adenovirus-mediated short hairpin RNA delivery system
复制标题
使用腺病毒介导的短发夹 RNA 递送系统显着抑制天布苏病毒包膜和 NS5 基因
DOI：
10.1016/j.meegid.2017.08.001
复制
发表时间：
2017-10-01
影响因子：
3.2
通讯作者：
Tang, Yi
中科院分区：
文献类型：
--
作者：
Wang, Hongzhi;Feng, Qiang;Tang, Yi
天布苏病毒 (TMUV) 是一种蚊媒黄病毒，于 20 世纪 70 年代首次在热带地区分离到。最近，我国鸭群中发现一种以卵巢出血和神经系统症状为特征的疾病，威胁家禽生产。然而，目前还没有合适的疫苗接种策略或有效的抗病毒药物来对抗 TMUV 感染。因此，迫切需要开发一种新的抗TMUV疗法。在这项研究中，我们报告了一种使用腺病毒载体系统抑制 TMUV 产生的有效短发夹 RNA (shRNA) 递送策略。使用基于TMUV的E和NS5蛋白基因专门设计的shRNA，载体表达的病毒基因、TMUV RNA复制和感染性病毒的产生在Vero细胞中不同程度地下调，其中shRNA（NS52）对抑制TMUV非常有效。使用人腺病毒5型shRNA递送系统，重组腺病毒（rAd-NS52）高效抑制TMUV增殖。此外，在TMUV感染的细胞中发现rAd-NS52诱导的病毒RNA拷贝的显着剂量依赖性抑制，这种抑制可以在感染后持续至少96小时。我们的结果表明，腺病毒介导的 shRNA 递送可以在未来的 TMUV 抗病毒治疗中发挥积极作用。
Tembusu virus (TMUV) is a mosquito-borne flavivirus, which was first isolated in the tropics during the 1970s. Recently, a disease characterized by ovarian haemorrhage and neurological symptoms was observed in ducks in China, which threatens poultry production. However, there is no suitable vaccination strategy or effective antiviral drugs to combat TMUV infections. Consequently, there is an urgent need to develop a new anti-TMUV therapy. In this study, we report an efficient short hairpin RNA (shRNA) delivery strategy for the inhibition of TMUV production using an adenovirus vector system. Using specifically designed shRNAs based on the E and NS5 protein genes of TMUV, the vector-expressed viral genes, TMUV RNA replication and infectious virus production were downregulated at different levels in Vero cells, where the shRNA (NS52) was highly effective in inhibiting TMUV. Using the human adenovirus type 5 shRNA delivery system, the recombinant adenovirus (rAd-NS52) inhibited TMUV multiplication with high efficiency. Furthermore, the significant dose-dependent inhibition of viral RNA copies induced by rAd-NS52 was found in TMUV-infected cells, which could last for at least 96 h post infection. Our results indicated that the adenovirus-mediated delivery of shRNAs could play an active role in future TMUV antiviral therapeutics.