Short hairpin RNAs targeting M and N genes reduce replication of porcine deltacoronavirus in ST cells
Porcine deltacoronavirus (PDCoV) is a recently identified coronavirus that causes intestinal diseases in neonatal piglets with diarrhea, vomiting, dehydration, and post-infection mortality of 50–100%. 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.
KeywordsPorcine deltacoronavirus RNA interference Short hairpin RNA Nucleocapsidgene Swine testicular cells
This research was supported by the Program of the Modern Agriculture Industry Technology System Foundation of Hebei Province (HBCT2018110207), Science and Technology Innovation Program of Hebei Province for graduate students (CXZZBS2019095), and Key R & D projects in Hebei Province, China (19226622D).
WYG performed the main experiments and wrote the paper; YL participated in editing the paper; BJL was involved in executing the study and editing the paper; YZZ and JHF participated in experimental design; JW, GFY, and SJC revised the paper.
Compliance with ethical standards
Conflict of interests
The authors declare that they have no conflicting interests.
This article does not contain any studies with animals performed by any of the authors.
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