NTCP Stable HepG2 Cell Line

The NTCP HepG2 cell line is a genetically engineered hepatocyte model developed to facilitate the study of Hepatitis B virus (HBV) infection and replication. This cell line was created by introducing the sodium taurocholate co-transporting polypeptide (NTCP) receptor into HepG2 cells via a lentivirus vector, which also expresses GFP for sorting purposes. The permissiveness of these cells to HBV infection is directly correlated with the levels of NTCP expression. Experiments showed that up to 50-60% of the HepG2/NTCP cells could be successfully infected with HBV, as determined by the presence of viral RNA transcripts and cytoplasmic HBV DNA. Additionally, Southern blot and PCR analyses confirmed the presence of covalently closed circular DNA (cccDNA) in infected cells. To enhance the utility of this cell line for advanced research, the CRISPR/Cas9 gene editing system was incorporated, allowing for precise targeting and cleavage of HBV cccDNA. This integration opens up new avenues for antiviral strategy development, as it demonstrates the potential to directly target and disrupt viral DNA within the nucleus. Overall, the NTCP HepG2 cell line serves as a powerful tool for investigating the molecular mechanisms of HBV infection and for testing novel antiviral therapies.