Transposon mouse models to elucidate the genetic mechanisms of hepatitis B viral induced hepatocellular carcinoma

Figure 6 Reverse genetic screening of individual HBV gene components using the Fah^-/-/SB^+/- transgenic mouse model. A: Construction of gene delivery expression vectors that carry various individual HBV gene-of-interest (GOI) components by PCR amplification and insertion of GOI into an entry clone (pENTR) and followed by LR clonase reaction (Life Technologies); B: Hydrodynamic tail vein injection of gene delivery expression vector into Fah^-/-/SB^+/- transgenic mouse; C: Liver tumors indicated by white arrowheads observed in Fah^-/-/SB^+/- mouse injected with a HBV gene component (left) after 160-d post-hydrodynamic injection. Fah^-/-/SB^+/- transgenic mouse injected with an empty gene delivery expression vector displayed normal liver morphology (right). EGFP: Enhanced green fluorescent protein gene; Luc: Luciferase gene; IR/DR: Inverted repeat/direct repeat sequences for Sleeping Beauty transposase binding and mobilization; pA: Polyadenalytion signal; IRES: Internal ribosome entry site; PGK: Phosphoglycerate kinase 1 promoter; CAG: Cytomegalovirus enhancer fused to the chicken β-actin promoter; attL/R/B sites: Recombination sites used by the Gateway cloning system (Life Technologies); ccdB: Topoisomerase poison from Escherichia coli. Scale bar: 0.5 cm.