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Title: Delayed Newcastle disease virus replication using RNA interference to target the nucleoprotein

Author
item HUTCHESON, JESSICA - University Of Georgia
item Susta, Leonardo
item STICE, STEVEN - University Of Georgia
item Afonso, Claudio
item WEST, FRANKLIN - University Of Georgia

Submitted to: Biologicals
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/26/2015
Publication Date: 6/4/2015
Publication URL: http://handle.nal.usda.gov/10113/61284
Citation: Hutcheson, J.M., Susta, L., Stice, S.L., Afonso, C.L., West, F.D. 2015. Delayed Newcastle disease virus replication using RNA interference to target the nucleoprotein. Biologicals. 43(4):274-280. doi: 10.1016/j.biologicals.2015.03.004.

Interpretive Summary: Newcastle disease virus (NDV) of very high virulence are rapidly spreading through Asia and the Middle East causing outbreaks of Newcastle disease (ND) characterized by significant illness and mortality in vaccinated poultry. These viruses represent a significant threat to the U.S. In isolated regions of developing countries where vaccine transport conditions are practically non-existing, new strategies to control the disease are needed. With the advent of stem cell technology it may be possible to generate animals that are resistant to ND. One promising strategy to generate disease resistant animals is to produce animals that express antisense nucleic acids that prevent virus replication. Here we have identified a possible target sequence that reduces virus replication in chicken cells.

Technical Abstract: Each year millions of chickens die from Newcastle disease virus (NDV) worldwide, leading to economic and food losses. Current vaccination campaigns have limitations including cost, administration, and thermostability. These problems are heightened in the developing world where constraints are more rigid, thus leading to interest in producing new antiviral strategies. RNA interference (RNAi) methodology is capable of inhibiting viral replication in vitro and in vivo. In this study, we utilize an RNAi strategy that produces microRNA (miRNA) targeting and knocking down NDVs nucleoprotein (NP), phosphoprotein (P), matrix (M), fusion (F), and large (L) proteins. NDV challenged chicken embryo fibroblast cells (DF-1) transfected with miRNA targeting NP, resulted in observable delay of NDV replication in culture as determined by increased cell survival. Subsequently transduced NP cells exposed to viral challenge demonstrated improved cell survival up to 48 hours post infection (h.p.i) and decreased viral growth in culture up to 72 h.p.i.