Inhibition of expression of rna polymerase with small interfering RNAs targeting a conserved motif in the respective viral genes in viruses of the family Flaviviridae.
The RNA-dependent RNA polymerase (RdRp) in viruses of the family Flaviviridae plays an important role in the viral replication process and in the forming of a replicase complex. We used small interfering RNAs (siRNAs) corresponding to the highly conservative Motif V of RdRp gene of different viruses to examine their role in modulating the expression of RdRp. Evaluation of the expression of RdRps was performed by the fluorescence, flow cytometry, Western blotting, and real-time PCR. We found that Classical swine fever virus (CSFV) siRNA could completely block the transcription and expression of RdRp. Additionally, Hepatitis C virus (HCV) siRNA could cause effective inhibition of RdRp, whereas Japanese encephalitis virus siRNA did not show significant repression of corresponding RdRp. These results demonstrated that siRNAs inhibited the expression of tested RdRps at the transcription level or at the posttranscriptional processing to a different extent.
Xu
Z
Z
College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, China.
Li
X
X
Liu
R
R
Si
Y
Y
Sun
M
M
Jin
M
M
Chen
H
H
Qian
P
P
D000818:Animals; D019677:Artificial Gene Fusion; D015153:Blotting, Western; D002460:Cell Line; D006692:Classical Swine Fever Virus / Q000235:genetics*; D017124:Conserved Sequence / Q000235:genetics; D004664:Encephalitis Virus, Japanese / Q000235:genetics*; D005434:Flow Cytometry; D015870:Gene Expression; D020868:Gene Silencing; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis; D012324:RNA-Dependent RNA Polymerase / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics
D034741:RNA, Small Interfering; D012367:RNA, Viral; D014764:Viral Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D012324:RNA-Dependent RNA Polymerase