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A novel recombinant cccDNA-based mouse model with long term maintenance of rcccDNA and antigenemia

Min Wu (邬敏), Cong Wang (王丛), Bisheng Shi (施碧胜), Zhong Fang (方钟), Boyin Qin (秦波音), Xiaohui Zhou (周晓辉), Xiaonan Zhang (张小楠), Zhenghong Yuan (袁正宏)
Shanghai Public Health Clinical Center, Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University


       The covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV) is critical for viral persistence in vivo. The lack of reliable, characterized and convenient small animal models for studying cccDNA persistence has long been a bottleneck for basic and translational research on HBV cure.A mouse model that can maintain intrahepatic cccDNA is urgently needed.   

         Through combining the Cre/loxP-mediated recombination and adeno-associated virus (AAV) vector delivery strategy, we establish a novel recombinant cccDNA (rcccDNA) mouse model (AAV-rcccDNA). AAV-rcccDNA mice supported long-term maintenance of intrahepatic rcccDNA. Quantitative PCR could detect the rcccDNA signal throughout the experiment duration (>51 weeks). Within 30 weeks after transduction, the rcccDNA in the liver of mice could be easily detected by Southern blotting. Furthermore, rcccDNA supported persistent serum antigenemia (>72 weeks) and intrahepatic surface and core antigen expression (>51 weeks). Flow cytometry analysis and single-cell RNA sequencing showed that AAV-rcccDNA mice displayed a compromised CD8+ T cell response. Meanwhile, minimal intrahepatic inflammation and fibrosis were observed. Furthermore, two anti-HBV compounds, AKEX0007, a post-transcriptional HBsAg suppressor and Bay 41-4109, a capsid allosteric modulator, were assessed in this AAV-rcccDNA mouse model. The changes of viral markers by these drugs were consistent with their mode of action although neither of them diminished the level of rcccDNA.

           Based on Cre/loxP-mediated rcccDNA production strategy, we adopted the AAV vector to establish a novel mouse model with long term maintenance of cccDNA and antigenemia. This mice model recapitulated the immune tolerant state of HBV infection, which will provide a suitable platform for studying cccDNA persistence and developing intervention strategies that would eventually break the tolerance and clear the virus.

Original paper

Image Collections

Disease phase
  • AAV-rcccDNA
  • AAV-rcccDNA antiviral treatment
Type of Staining
  • HBsAg(brown) hematoxylin
  • HBcAg(brown)-hematoxylin
Case No.