Differential inhibition of lamivudine-resistant hepatitis B virus by allele-specific RNAi.
Drug-resistant hepatitis B virus (HBV) is a serious problem affecting antiviral therapy. In this study, two long-term eukaryotic cell lines with full-length HBV were constructed and contained either lamivudine-resistant mutants (HBV-YIDD) or wild-type virus (HBV-wt). High levels of intracellular viral DNA replication were observed continuously after transfecting the plasmids into HepG2 cells, and HBV surface antigen (HBsAg) and HBV e antigen (HBeAg) were secreted into the cell culture supernatant. A series of experiments showed differential inhibition of HBV gene expression and replication by four specific siRNAs, according to the principles of allele-specific RNAi technology. The results showed that the designed siRNAs with a mismatch in the sixteenth nucleotide of the guide strands could effectively discriminate the HBV-YIDD mutants from the wild-type alleles, thus providing a new insight into the development of antiviral therapy with differing or complementary patterns characteristic of lamivudine-resistant HBV.
Teng
Xu
X
Department of Microbiology, Harbin Medical University, Nangang District, Harbin, Heilongjing Province, China.
Xu
Wei-Zhen
WZ
Hao
Mei-Li
ML
Fang
Yong
Y
Zhao
Ying-Xin
YX
Chen
Si-Jia
SJ
Li
Di
D
Gu
Hong-Xi
HX
D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry; D024882:Drug Resistance, Viral; D020868:Gene Silencing; D005814:Genes, Viral; D006514:Hepatitis B Surface Antigens / Q000378:metabolism; D006513:Hepatitis B e Antigens / Q000378:metabolism; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics*; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D003470:Culture Media; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D034741:RNA, Small Interfering; D019259:Lamivudine