[Effect of short interfering RNAs on hepatitis C Virus 5' untranslated region expression].
OBJECTIVE
To evaluate interfering effect of several short interfering RNAs (siRNA) on HCV 5' untranslated region(5' UTR).


METHODS
The green fluorescent protein (GFP) was used as reporter gene. A fused gene of HCV-5'UTR and GFP was constructed. It was cloned into the plasmid pCDNA3.1 named as pcDNA-HCV-5'UTR-GFP. Three siRNAs were designed and transfected into HepG2 cells with pcDNA-HCV-5'UTR-GFP. The change of the fluorescence intensity of HepG2 cells was shown by fluorescence microscopy and numerically detected under 488 nm wave length by flow cytometry.


RESULTS
The fused gene of HCV-5'UTR and GFP was successfully constructed. The seven groups displayed inhibitory effects on the gene expression of GFP. The inhibition rates of siRNA A, B and C were 68.4 percent, 72.6 percent and 75.6 percent, respectively. The inhibitory rates of siRN A + B, siRN B +C and siRN A +C were 91.8 percent, 87.2 percent and 92.4 percent, respectively. The inhibitory rates of siRN A+B +C was the highest, up to 95.7 percent.


CONCLUSIONS
These siRNAs could inhibit expression of HCV 5'UTR gene, the inhibitory effect of combined siRNA was better than that of single siRNAs.
Chen
Zhao-jun
ZJ
Clinical Laboratory Center, Hangzhou Second Hospital, Hangzhou 310015, China. hzczj2006@163.com
Zhang
La-hong
LH
Li
Wei
W
Liu
Xia
X
D020121:5' Untranslated Regions / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
D020121:5' Untranslated Regions; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins