Transgenome transcription and replication in the liver and extrahepatic tissues of a human hepatitis B virus transgenic mouse.

We have produced a transgenic mouse (B32-1) carrying the complete genome of the human hepatitis B virus (HBV). High titers of the viral surface (HBsAg) and the e antigen (HBeAg) were detected in the serum of the mouse. In the liver and 12 of 16 extrahepatic tissues analyzed, Northern blot hybridization indicated the presence of the 2.1-kilobase (kb) and the 3.5-kb major HBV transcripts. A liver cDNA library was constructed from which the liver RNAs from four cDNA clones with splicing were found. Sequencing analysis showed that the splicing occurred between nucleotides 2451 and 487 of the viral genome, resulting in a truncated viral polymerase gene, as in human hepatocytes. Southern blot analysis of total DNA preparations of the tissues revealed the presence of episomal HBV genome, indicating replication of the viral transgenome in these tissues. However, replication was detected only in some but not all of the tissues that transcribed the 3.5-kb RNA. Partial double-stranded as well as full-length and subgenomic-length single-stranded HBV DNA species of discrete sizes were detected which may represent replication intermediates of preferred replication termination sites of the HBV transgenome. Since many molecular characteristics of mouse B32-1 were similar to those found in HBV-infected humans, HBV transgenic mice similar to B32-1 would be useful in further elucidation of other aspects of the replication and transcription mechanisms of HBV in the liver and extrahepatic tissues.