[Transcription of antisense RNA of hepatitis B virus through retroviral mediated gene transfer].

To probe the ways by which the antisense gene of hepatitis B virus (HBV) can be transferred and transcripted in eukaryotic cells to inhibit HBV replication and expression. Two retroviral vectors that carried antisense gene of hepatitis B virus (HBV) PreC/C or PreS/S region were constructed. The HBV ayw PreC/C and PreS/S fragments were inserted into the vector pDO. R cloning site in the sense or antisense orientation and the recombinant retroviral vectors were then transfected into PA317 packaging cells by calcium phosphate coprecipitation, respectively. The stably transformed G418-resistant PA317 cells were selected and the recombinant retroviruses released from transfected G418-resistant PA317 cells were assayed by G418 selection method, using NIH 3T3 cells as target cells. Southern blot and RNA Dot blot analysis showed that the recombinant retroviral vector sequences were stably integrated into the chromosome of transfected PA317 cells and the antisense RNA of HBV PreS/S or PreC/C gene also transcripted in the transduced NIH 3T3 cells. These results suggested that antisense gene of HBV can be successfully transferred and transcripted in target cells through retroviral vector mediated gene transfer and the antisense retroviral vectors may be potentially useful for anti-HBV gene therapy.