Specific interference shRNA-expressing plasmids inhibit Hantaan virus infection in vitro and in vivo

• Published in: Acta pharmacologica Sinica Vol. 37; no. 4; pp. 497 - 504
• Main Authors: Liu, Yuan-yuan, Chen, Liang-jun, Zhong, Yan, Shen, Meng-xin, Ma, Nian, Liu, Bing-yu, Luo, Fan, Hou, Wei, Yang, Zhan-qiu, Xiong, Hai-rong
• Format: Journal Article
• Language: English
• Published: United States Nature Publishing Group 01.04.2016
• Subjects: Animals; Cercopithecus aethiops; Hantaan virus; Hantaan virus - physiology; Hemorrhagic Fever with Renal Syndrome - genetics; Hemorrhagic Fever with Renal Syndrome - therapy; Hemorrhagic Fever with Renal Syndrome - virology; Mice, Inbred BALB C; Original; Plasmids; RNA, Small Interfering - genetics; RNA干扰; shRNA; Vero Cells; Virus Replication; 体内; 体外; 抗病毒作用; 汉坦病毒; 病毒感染; 质粒转染
• ISSN: 1671-4083; 1745-7254
• DOI: 10.1038/aps.2015.165

Aim： To investigate the antiviral effects of vectors expressing specific short hairpin RNAs （shRNAs） against Hantaan virus （HTNV） infection in vitro and in vivo. Methods： Based on the effects of 4 shRNAs targeting different regions of HTNV genomic RNA on viral replication, the most effective RNA interference fragments of the S and M genes were constructed in pSilencer-3.0-H1 vectors, and designated pSilencer-S and pSilencer-M, respectively. The antiviral effect of pSilencer-S/M against HTNV was evaluated in both HTNV-infected Vero-E6 cells and mice. Results： In HTNV-infected Vero-E6 cells, pSilencer-S and pSilencer-M targeted the viral nucleocapsid proteins and envelope glycoproteins, respectively, as revealed in the immunofluorescence assay. Transfection with pSilencer-S or pSilencer-M （1, 2, and 4 pg） markedly inhibited the viral antigen expression in dose- and time-dependent manners. Transfection with either plasmid （2 pg） significantly decreased HTNV-RNA level at 3 day postinfectin （dpi） and the progeny virus titer at 5 dpi. In mice infected with lethal doses of HTNV, intraperitoneal injection of pSilencer-S or pSilencer-M （30 pg） considerably increased the survival rates and mean time to death, and significantly reduced the mean virus yields and viral RNA level, and alleviated virus-induced pathological lesions in lungs, brains and kidneys. Conclusion： Plasmid-based shRNAs potently inhibit HTNV replication in vitro and in vivo. Our results provide a basis for development of shRNA as therapeutics for HTNV infections in humans.