Abstract
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 UL42 gene of human herpes simplex virus type 1 (HSV-1), which encodes a multifunctional polypeptide that is vital for virus DNA replication, binding to DNA, stably associating with the virus DNA polymerase (Pol), and acting to increase the length of DNA chains synthesized by Pol. HeLa cell line was used for HSV 1 infection and SiRNA transfection was done to suppress UL-42 gene in cell culture. The decrease in titer of HSV 1 was observed by rReal Time PCR to detect the drop in HSV 1 DNA synthesis and translation. The inhibition rates of siRNA1 and siRNA2 on HSV-1 plaque formation were reported and Comparing with virus control, siRNA1 and siRNA2 reduced DNA replication HSV-1. The decision whether the decrease in the number of HSV-1 plaques was due to siRNA silencing expression of the UL42 gene, a real-time PCR indicating that UL42-specific siRNAs blocked the expression of the UL42 gene significantly. Comparing with virus control, siRNA1 and siRNA2 reduced the expression of UL42 gene. In this study the synthetic siRNA silenced UL42 mRNA expression effectively and specifically and inhibited HSV-1 replication and also our data offer new possibilities for RNAi as a genetic tool for inhibition of HSV-1 replication.
Highlights
Herpes simplex virus type 1 (HSV-1) is a large, nuclearreplicating, dsDNA virus which is the member of the alpha herpes virus subfamily
We mainly focused on UL42 gene of HSV-1 and designed only one small interfering RNA molecule that targets the mRNA of the early gene UL42 as a possible therapeutic strategy in herpes simplex virus (HSV) infections and the activity of HSV-1 in vitro evaluated after challenging with siRNA therapy
HeLa cells were transfected with UL42-specific siRNAs or negative control siN.C and infected with 25 PFU of HSV-1
Summary
Herpes simplex virus type 1 (HSV-1) is a large, nuclearreplicating, dsDNA virus which is the member of the alpha herpes virus subfamily. Herpes virus type 1 is the most common cause of cold sores around the mouth and lips (fever blisters). This virus is transmitted through oral secretions or wounds on the skin can be spread through kissing or sharing objects such as toothbrushes or eating utensils. There are three kinetic groups of proteins playing a role in herpes simplex virus replication: immediate early, early, and late. The seven of early (β) genes, DNA polymerase (UL30), DNA binding protein (UL42 and UL29/ ICP8), ORI binding protein (UL9), and the helicase/primase complex (UL5, UL8 and UL52) have been shown to be essential in virus replication, while the rest of the (β) gene products play only a partial role. In addition to DNA polymerase activity, the U 30 acts as a DNAdependent polynucleotide synthesis, 3’-5’ exoLnuclease and RNase H [7,8]
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