Establishment of a screening system for selection of siRNA target sites directed against hepatitis B virus surface gene

	Establishment of a screening system for selection of siRNA target sites directed against hepatitis B virus surface gene
	Zhou, XM; Lin, JS; Shi, Y; Tian, DA; Huang, HJ; Zhou, HJ; Jin, YX
刊名	ACTA BIOCHIMICA ET BIOPHYSICA SINICA
	2005
卷号	37 期号:5 页码:310-316
关键词	RNA interference (RNAi) small interference RNA (siRNA) hepatitis B virus surface gene (HBs gene)
通讯作者	Jin, YX (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, Shanghai 200031, Peoples R China.,yxjin@sibs.ac.cn
英文摘要	To study the inhibitory effects of plasmid-derived small interfering RNA (siRNA) and synthetic siRNA on the expression of the hepatitis B virus surface (HBs) gene, three plasmid-derived siRNAs and one synthetic siRNA that complement the coding region of the HBs gene were prepared. The HBs expression plasmid pHBs-EGFP was also constructed. HeLa cells were co-transfected with pHBs-EGFP and the above siRNAs. The HBs mRNA quantities were measured by reverse-transcription PCR, and the level of HBs-EGFP fusion protein was quantified by fluorescent microscope. The concentrations of the hepatitis B virus surface antigen (HBsAg) derived from the culture supernatant of transfected HepG2.2.15 cells were measured by an enzyme-linked immunosorbent assay (ELISA) kit. The results showed that the three plasmid-derived siRNAs and the synthetic siRNA can effectively reduce the quantities of HBs mRNA and protein. The plasmid-derived siRNA psiRNA1 was found to be the most effective inhibitor of HBs expression. It can inhibit HBs-EGFP expression by 63.3% and suppress HBs mRNA by 75.6%. To further substantiate the above observations, psiRNA1 was transfected into HepG2.2.15 cells (an HBV secreting cell line). The transfections resulted in almost complete blockage of HBsAg production, whereas control vector-transfected cells secreted high levels of HBsAg 7 days post-transfection. In conclusion, our data suggests that RNA interference (RNAi) is an efficient approach for reducing the level of HBs transcripts and proteins and for suppressing HBsAg production.
学科主题	Biochemistry & Molecular Biology; Biophysics
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	RNA INTERFERENCE ; IN-VIVO ; INHIBITION ; DESIGN ; MICE ; DNA ; REPLICATION ; LAMIVUDINE ; EXPRESSION ; REGION
收录类别	SCI
语种	英语
WOS记录号	WOS:000229304400004
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1915]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Zhou, XM,Lin, JS,Shi, Y,et al. Establishment of a screening system for selection of siRNA target sites directed against hepatitis B virus surface gene[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2005,37(5):310-316.
APA	Zhou, XM.,Lin, JS.,Shi, Y.,Tian, DA.,Huang, HJ.,...&Jin, YX.(2005).Establishment of a screening system for selection of siRNA target sites directed against hepatitis B virus surface gene.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,37(5),310-316.
MLA	Zhou, XM,et al."Establishment of a screening system for selection of siRNA target sites directed against hepatitis B virus surface gene".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 37.5(2005):310-316.