Inhibition of HBs-GFP fusion gene expression by RNA interference / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 110-115, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-353236
ABSTRACT
<p><b>OBJECTIVE</b>To develop an effective report gene system to test the effect of small interfering RNA (siRNA).</p><p><b>METHODS</b>HBV S gene was fused with enhanced green fluorescent protein (EGFP) gene to form HBs-GFP and the plasmid containing HBs-GFP was constructed. A vector expressing small hairpin RNA (shRNA) pAVU6 + 4sh357 was also constructed. Two plasmids were co-transfected into HepG2 cells transiently. The fluorescence of HBs-GFP was detected by fluorescence-activated cell sorting (FACS). The mRNA expression in HepG2 cells was detected by conventional RT-PCR and real-time PCR.</p><p><b>RESULTS</b>siRNA inhibited the expression of HBs-GFP 72 hours post transfection. The fluorescence of HBs-GFP in HepG2 cells treated with pAVU6+4sh357 was reduced by 55.4% compared with that of controls. The HBs-GFP expression in HepG2 cells treated with pAVU6+4sh357 was reduced by 76.3% and 90% as measured with conventional RT-PCR and real-time PCR, respectively.</p><p><b>CONCLUSION</b>This investigation demonstrated siRNA derived from shRNA expression vectors can inhibit the expression of HBs-GFP in HepG2 cells.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Recombinant Fusion Proteins
/
Tumor Cells, Cultured
/
Transfection
/
Gene Expression Regulation, Viral
/
Hepatitis B virus
/
Carcinoma, Hepatocellular
/
RNA, Small Interfering
/
RNA Interference
/
Green Fluorescent Proteins
Limits:
Humans
Language:
Chinese
Journal:
Journal of Zhejiang University. Medical sciences
Year:
2005
Type:
Article
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