[SiRNA targeting ICP4 attenuates HSV-1 replication].
HSV-1, a neurotropic virus, always leads to severe nervous symptoms. It is hard to completely eradicate the latent viruses after conventional therapy so that recurrence is inevitable. ICP is a key regulator for HSV replication and transcription that determines the cytolytic infection or latent state. In search of new anti-virus strategy targeting HSV-1ICP4, two pairs of siRNA were designed, and a recombinant eukaryotic lentiviral expression plasmid pLKO-puro(r)-hU6-siRNA was constructed. Vero cells were transfected with the designed siRNAs by Lipofectamine 2000 and four stable monoclonal cell lines were established by puromycin screening method. The ICP4 expression at mRNA level was detected with real-time PCR, and the HSV-1 replication was measured with TCID50 assay. SiRNA was shown as an effective way to inhibit the expression of ICP4 in monoclonal cell lines. Meanwhile, HSV-1 replication was significantly inhibited when ICP4 was shut down by siRNA. We conclude that siRNA targeting ICP4 attenuates HSV-1 replication. Further more, multi-site siRNAs show stronger inhibitory effect on viral replication, which may be an effective and feasible approach for biological anti-viral drugs.
Liu
Yu-tao
YT
Department of Neurology, The First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China. hnliuyt@163.com
Song
Bo
B
Wang
Ya-lun
YL
Xu
Yu-ming
YM
Han
Zhi-qiang
ZQ
Zhao
Xin-yu
XY
Jia
Li-jie
LJ
D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000502:physiology*; D006801:Humans; D017874:Immediate-Early Proteins / Q000172:deficiency* / Q000235:genetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D014709:Vero Cells; D014779:Virus Replication / Q000235:genetics*
D017874:Immediate-Early Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C048925:herpes simplex virus, type 1 protein ICP4