Screen To Identify The Antiviral Proteins And Explore The Effect Of Prostaglandin Receptor

The innate immunity is the first line of defense against microbial infection,which is related to infectious and noninfectious disease.Among them,the type-1 interferon(IFN-?/?)is an important cellular factors which can mediate the antiviral activity.The type-1 interferon stimulate the IFNR(interferon receptor)-JAK/STAT signal pathway and transcriptionally regulate hundreds of interferon-stimulated genes(ISGs).Collectively,they can target almost any step in a virus life cycle(e.g.entry,uncoating,genome replication,particle assembly,egress)and influence the growth and survival of the host cell.Consequently,the type-1 IFN help the cell defense against multiple microbial infection.Although hundreds of ISGs have been identified more than 25 years ago,only a few of them have been characterized with respect to antiviral effect.For most ISG products,little is known of their antiviral activity,target specificity and mechanisms of action.Herpes simplex virus type 1(HSV-1)is a double stranded DNA virus.It is a widespread virus and it is infects approximately 80%of adults worldwide.HSV-1 infection can cause the fatal childhood HSV-1 encephalitis(HSE).To conduct the screen we use the RNA deep sequencing to identify the gene which can be induced by interferon in the HEK293 cell,and construct an almost 600 plasmid pool.These ISGs were tested for their ability to inhibit the replication of two important human and animal viruses HSV and VSV.Using this screen we find more than 10 candidate.Through the gene editing method based on CRISPR-CAS9 nuclease we identify the prostaglandin 12 receptor(IP)and the same family member prostaglandin D2 receptor(DP)have the antiviral activity.Using the overexpress and CRISPR-CAS9 nuclease we can confirm that the downstream factor G?q has the important role in the antiviral activity.In addition overexpressing Gas protein and the downstream factor PKAG can enhance the effect of Gaq respectively.These results may provide a new perspective for the exploration of pharmaceutical approaches against HSV-1 related disease.