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Expression of the hepatitis C virus genome in rat liver after cationic liposome‐mediated in vivo gene transfer
Author(s) -
Takehara Tetsuo,
Hayashi Norio,
Miyamoto Yasuhide,
Yamamoto Masato,
Mita Eiji,
Fusamoto Hideyuki,
Kamada Takenobu
Publication year - 1995
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840210323
Subject(s) - in vivo , biology , microbiology and biotechnology , hepatitis c virus , genetic enhancement , complementary dna , gene expression , cationic liposome , gene , transfection , virology , virus , biochemistry , genetics
The lack of a small animal model of hepatitis C virus (HCV) infection has impeded elucidation of the pathogenesis of HCV. The aim of this study was to develop an HCV‐expressing animal model by means of cationic liposome‐mediated in vivo gene transfer. To examine the feasibility of this strategy, pActLacZ, an expression vector composed of the LacZ gene driven by the β‐actin promoter, complexed with lipofectin, 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, but not in biliary epithelial cells. Maximal expression was observed at a DNA to lipofectin ratio of 1:4. Based on this observation, pAGS3M091, an expression vector containing the full length of HCV complementary DNA (cDNA) preceded by the β‐actin promoter, was evaluated. Two days after in vivo intrabiliary administration of pAGS3M091 complexed with lipofectin, polymerase chain reaction (PCR) amplification of reverse‐transcribed liver RNA demonstrated the 5′ and 3′ portions of HCV transcripts derived from pAGS3M091. Immunohistochemical analysis showed the HCV core protein in a small number of hepatocytes scattered in the hepatic lobules. We conclude that the full‐length HCV genome was successfully expressed in adult rat liver by means of cationic liposome‐mediated in vivo gene transfer. This model will be useful for detemining the immunopathological role of HCV in vivo .