RNA interference protects horse cells in vitro from infection with Equine Arteritis Virus

Equine Arteritis Virus (EAV) belongs to the Arteriviridae and causes viral arteritis in horses. In an attempt to develop novel and save therapies against the infection it was tested whether EAV is susceptible to RNA interference (RNAi) in an equine in vitro system. Horse cells were transfected with chemically synthesized small interfering RNA oligonucleotides (siRNAs) and challenged with EAV. Application of these siRNAs led to a significant protection of the cells, and virus titers decreased drastically. siRNAs derived from DNA plasmids expressing small hairpin RNAs (shRNAs) were also effective. The protection was most pronounced with two siRNAs targeting the open reading frame 1 (coding for non-structural proteins), whereas siRNAs targeting sequences for several structural proteins had less or no effect. In addition, it was investigated whether RNAi could be used to treat cells with an already established viral infection. Only application of the siRNAs shortly after viral challenge led to significant survival rates of the cells, whereas transfection at later time points caused much less benefit for the cells. These findings are discussed in a perspective of using RNAi as a therapeutic approach to combat EAV.