Silencing of the entry fusion complex genes of amsacta moorei entomopoxvirus (AMEV)

Amsacta moorei entomopoxvirus (AMEV) has become an important research model for elucidating the molecular mechanisms of vertebrate-infecting poxviruses, as it naturally infects invertebrates. In addition, AMEV is a significant insect virus with potential applications as an expression vector, a gene therapy vector, and a microbiological control agent against agriculturally important pests. Whereas the cell entry-fusion complex (EFC) of vaccinia virus is composed of 11 viral proteins, it has been proposed that only 9 genes products constitute the EFC in AMEV. The aim of this study was to investigate the effects of silencing AMEV EFC genes on the production of new viruses. Using RNA interference (RNAi), long double-stranded RNAs (dsRNAs) specific to AMEV EFC genes were synthesized. According to the transfection-infection results, the titer of newly produced virus in the RNAi application was detected by GFP fluorescence, and a reduction of virüs titer was determined for many genes. The decrease in virus titer was detected as 64.98-fold for dsAMV186-Am∆sph/gfp and 32.93-fold for dsAMV232-Am∆sph/gfp applied alone, while it was detected as 12.68-fold for dsAMV217/dsAMV243-Am∆sph/gfp and 12.35-fold for dsAMV232/dsAMV243-Am∆sph/gfp applied together.