In vivo transfection of rat liver with hepatitis C virus cDNA using cationic liposome-mediated gene delivery.
The lack of a small animal model for hepatitis C virus (HCV) infection has impeded elucidation of the pathogenesis of this virus. The aim of this study was to develop an HCV-expressing animal model using cationic liposome-mediated in vivo gene transfer. To examine the feasibility of this strategy, an expression vector composed of the LacZ gene driven by the beta-actin promoter, pActLacZ, was injected retrogradely into the common bile ducts of adult rats. X-Gal histochemical staining clearly showed that the LacZ gene was expressed in hepatocytes. Maximal expression was observed at a DNA:lipofectin ratio of 1:4. Based on this observation, an expression vector containing the full-length of HCV cDNA, pAGS3M091, was evaluated in adult rats. Two days after intrabiliary administration of pAGS3M091, PCR amplification of reverse-transcribed liver RNA demonstrated the 5' and 3' portions of HCV transcripts derived from pAGS3M091. Immunohistochemical analysis revealed the HCV core protein in a small number of hepatocytes scattered in the lobules. Thus, the full-length of the HCV genome was successfully expressed in adult rat liver using liposome-mediated in vivo gene transfer.
Hayashi
N
N
First Department of Medicine, Osaka University School of Medicine, Japan.
Takehara
T
T
Kamada
T
T
D000818:Animals; D018076:DNA, Complementary / Q000235:genetics*; D018014:Gene Transfer Techniques; D016174:Hepacivirus / Q000235:genetics*; D008081:Liposomes; D008099:Liver / Q000821:virology*; D051381:Rats; D014162:Transfection
D018076:DNA, Complementary; D008081:Liposomes