[Experimental study of inhibition of hepatitis B by dual-target antisense RNA].
OBJECTIVE
To investigate the effect of retroviral vector expressing the dual--target antisense RNA of hepatitis B virus (HBV) on the replication and expression of HBV.


METHODS
Recombinant retroviral vector plasmids expressing dual-target antisense RNA complementary to HBV X (1400 - 1430) and P (2375 - 2405) were constructed and transduced into 2.2.15 cells. The experimental cells were divided into PLXSN + X, PLXSN + P, PLXSN + X/P, and PLXSN + Xpos/P pos groups. The HBV expression was tested by ELISA method. HBV DNA and RNA were tested by FQ-PCR.


RESULTS
The inhibition rates of 2.2.15 cells transduced with recombinant vector plasmids (PLXSN + X group, PLXSN + P group, especially the PLXSN + X/P group) on expression of HBV DNA and RNA were higher than those of blank control group, PLXSN group, and PLXSN + Xpos/Ppos group. The expression of the three ORFs, S, C, and P were significantly inhibited.


CONCLUSIONS
The expression of HBV antisense RNA in 2.2.15 cells can inhibit the replication and expression of HBV. The inhibitory effect of dual antisense-RNA group is higher than that of single antisense-RNA group. Dual antisense RNA has the potentiality in anti-HBV gene therapy.
Wu
C
C
Department of Infectious Diseases, First Hospital, Peking University, Beijing 100034, China.
Zeng
Z
Z
Wang
Q
Q
D016475:3T3 Cells; D000818:Animals; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics / Q000254:growth & development*; D006801:Humans; D051379:Mice; D016372:RNA, Antisense / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured; D054334:Viral Regulatory and Accessory Proteins; D014779:Virus Replication / Q000235:genetics*
D004279:DNA, Viral; D016372:RNA, Antisense; D012367:RNA, Viral; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein