RESUMO
RNA interference (RNAi) is a process by which introduced small interfering RNA (siRNA) can cause the specific degradation of mRNA with identical sequences. In this study, we applied siRNAs targeting the UL39 gene of human herpes simplex virus type 1 (HSV-1), which encodes the large subunit of ribonucleotide reductase, ICP6. Using an ICP6 expression reporter plasmid and an in vitro model of infection, we showed that synthetic siRNA silenced effectively and specifically UL39 mRNA expression and inhibited HSV-1 replication. Our work offers new possibilities for RNAi as a genetic tool for inhibition of HSV-1 replication.
Assuntos
Herpesvirus Humano 1/crescimento & desenvolvimento , RNA Interferente Pequeno/genética , Ensaio de Placa Viral , Proteínas Virais/biossíntese , Animais , Chlorocebus aethiops , Inativação Gênica , Herpesvirus Humano 1/genética , RNA Mensageiro/genética , RNA Viral/genética , Células Vero , Proteínas Virais/genética , Replicação ViralRESUMO
To explore the anti-HSV-1 effect of silencing gD gene expression by RNA interference, five 21-nucleotide duplex small interfering RNAs(siRNAs) targeting the HSV1 gD sequence were designed and the gD-EGFP fusion gene expression vector was constructed, then co-transfected into Vero cell, and screened the effective siRNA through analyzing the intensity of the EGFP fluorescence. Finally, the anti-HSV1 effect was confirmed by plaque reduction assay, real-time PCR and daughter virus titration of HSV1 infected Vero cells transfected with siRNAs. The study demonstrated that siRNAs could effectively and specifically inhibit gD gene expression in HSV1-infected cells, but only had a little effect on HSV1 infection, so taking gD as the target of siRNA against HSV1 needs further study.
