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.
Artigo
em Chinês
| 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>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Patologia
/
Proteínas Recombinantes de Fusão
/
Células Tumorais Cultivadas
/
Transfecção
/
Regulação Viral da Expressão Gênica
/
Vírus da Hepatite B
/
Carcinoma Hepatocelular
/
RNA Interferente Pequeno
/
Interferência de RNA
/
Proteínas de Fluorescência Verde
Limite:
Humanos
Idioma:
Chinês
Revista:
Journal of Zhejiang University. Medical sciences
Ano de publicação:
2005
Tipo de documento:
Artigo
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