Open AccessNovel reporter system to monitor early stages of the hepatitis B virus life cycle
Author(s) -
Nishitsuji Hironori,
Ujino Saneyuki,
Shimizu Yuko,
Harada Keisuke,
Zhang Jing,
Sugiyama Masaya,
Mizokami Masashi,
Shimotohno Kunitada
Publication year - 2015
Publication title -
cancer science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 141
eISSN - 1349-7006
pISSN - 1347-9032
DOI - 10.1111/cas.12799
Subject(s) - hepatitis b virus , virology , plasmid , recombinant dna , virus , biology , cell culture , reporter gene , viral life cycle , hepatitis b virus pre beta , transfection , hepatocyte , gene , hepatitis b , microbiology and biotechnology , gene expression , hepatitis b virus dna polymerase , viral replication , in vitro , genetics
A recombinant hepatitis B virus (HBV) expressing NanoLuc (NL) (HBV/NL) was produced by cotransfecting a plasmid containing a 1.2‐fold HBV genome carrying the NL gene with a plasmid bearing a packaging‐defective 1.2‐fold HBV genome into a human hepatoma cell line, HepG2. We found that NL activity in HBV/NL‐infected primary hepatocytes or sodium taurocholate cotransporting polypeptide‐transduced human hepatocyte‐derived cell lines increased linearly for several days after infection and was concordant with HBV RNA levels in the cells. Treatment of the virus‐infected cells with HBV inhibitors reduced NL activity in a dose‐dependent manner. Detection of HBV/NL infection, monitored by NL activity, was highly sensitive and less expensive than detection using the conventional method to evaluate HBV infection. In addition, because we also studied host factors, this system is applicable not only for studying the HBV life cycle, but also for exploring agent(s) that regulate HBV proliferation.