๐ Short hairpin RNAs targeting M and N genes reduce replication of porcine deltacoronavirus in ST cells
ยฉ 2019, Springer Science+Business Media, LLC, part of Springer Nature. Porcine deltacoronavirus (PDCoV) is a recently identified coronavirus that causes intestinal diseases in neonatal piglets with diarrhea, vomiting, dehydration, and post-infection mortality of 50โ100%. Currently, there are no effective treatments or vaccines available to control PDCoV. To study the potential of RNA interference (RNAi) as a strategy against PDCoV infection, two short hairpin RNA (shRNA)-expressing plasmids (pGenesil-M and pGenesil-N) that targeted the M and N genes of PDCoV were constructed and transfected separately into swine testicular (ST) cells, which were then infected with PDCoV strain HB-BD. The potential of the plasmids to inhibit PDCoV replication was evaluated by cytopathic effect, virus titers, and real-time quantitative RT-PCR assay. The cytopathogenicity assays demonstrated that pGenesil-M and pGenesil-N protected ST cells against pathological changes with high specificity and efficacy. The 50% tissue culture infective dose showed that the PDCoV titers in ST cells treated with pGenesil-M and pGenesil-N were reduced 13.2- and 32.4-fold, respectively. Real-time quantitative RT-PCR also confirmed that the amount of viral RNA in cell cultures pre-transfected with pGenesil-M and pGenesil-N was reduced by 45.8 and 56.1%, respectively. This is believed to be the first report to show that shRNAs targeting the M and N genes of PDCoV exert antiviral effects in vitro, which suggests that RNAi is a promising new strategy against PDCoV infection.
keywords
๐ cytopathic effect (91)
๐ tissue culture (67)
๐ first report (99)
๐ cell culture (240)
author
๐ค Gu, Wen yuan
๐ค Li, Yan
๐ค Liu, Bao jing
๐ค Wang, Jing
๐ค Yuan, Guang fu
๐ค Chen, Shao jie
๐ค Zuo, Yu Zhu
๐ค Fan, Jing Hui
year
โฐ 2019
journal
๐ Virus Genes
issn
๐ 1572994X 09208569
volume
55
number
6
page
795-801
citedbycount
0
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