[Construction of plasmid stably expressing shRNA and inhibition of HBV replication and expression].
OBJECTIVE
To construct the vector stably expressing small hair RNA(shRNA) and investigate the effect of shRNA expressed with DNA vector on HBV replication and expression.


METHODS
The vector expressing shRNA designated pU6 was constructed by U6 promoter. The sequence of shRNA targeting on specific region of HBV genome against HBV was designed and synthesized, then cloned into pU6. The plasmid designated pU6B/HBVi was transfected into 2.2.15 cells using Lipofectamine 2000 reagent. The HBsAg and HBeAg in the supernatants of the transfected cells were measured by ELISA. The HBV copies in the transfected cells were measured by quantitative PCR.


RESULTS
Compared with controls, not only concentration of HBsAg and HBeAg but also HBV copies in transfected 2.2.15 cells decreased significantly by the plasmid expressing shRNA against HBV.


CONCLUSIONS
HBV replication and expression in 2.2.15 cells are inhibited by stably expressed shRNA.
Zhang
Hong-Bin
HB
Department of Medical Research, Guangzhou Army General Hospital, Guangzhou, China. zhangwater@tom.com
Wu
Jie
J
Zheng
Wen-Ling
WL
Ma
Wen-Li
WL
Xian
Jiang
J
Xie
Wen-Yue
WY
Li
Wei
W
Wang
Jie
J
D000818:Animals; D002460:Cell Line; D017077:Culture Media, Conditioned / Q000378:metabolism; D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D055029:Inverted Repeat Sequences; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
D017077:Culture Media, Conditioned; D006514:Hepatitis B Surface Antigens; D034741:RNA, Small Interfering