Efficient inhibition of hepatitis B virus replication by small interfering RNAs targeted to the viral X gene in mice.
The hepatitis B virus (HBV), as a major cause of acute and chronic hepatitis in humans, contains a partial double-stranded circular DNA genome of 3.2kb that is transcribed into the 3.5-, 2.4-, 2.1-, and 0.7-kb viral transcripts by the host RNA polymerase II. The HBV X (HBx) gene is consistently expressed in all four HBV viral mRNAs and thus an ideal target for developing viral inhibitors via a gene therapeutic approach. In this study, we show that two HBx-specific small interfering RNAs (siRNA), HBx1 and HBx3, significantly decrease both viral RNA and protein levels, and completely block replication in cultured cells co-transfected with a siRNA expression plasmid and an HBV replication-competent vector. To further confirm these antiviral activities of selected siRNAs in small animals, we established acute and chronic HBV mouse models by hydrodynamic injection of this plasmid containing the full-length HBV genome. Selected HBx-specific siRNAs also induced a significant anti-viral effect in living animals. Our findings should facilitate the development of an alternative therapeutic agent against HBV infection, particularly HBV-derived hepatocellular carcinoma (HCC) in which HBx has been known as one of the major pathological factors.
Shin
Duckhyang
D
Immunology and Virology Group, Mogam Biotechnology Research Institute, 341 Pojung-ri, Guseong-eup, Yongin-city, Kyonggi-do 449-913, South Korea.
Kim
Soo In
SI
Kim
Meehyein
M
Park
Mahnhoon
M
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015316:Genetic Therapy; D006509:Hepatitis B / Q000628:therapy* / Q000821:virology; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D034622:RNA Interference; D012333:RNA, Messenger / Q000096:biosynthesis; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics*; D012367:RNA, Viral / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators / Q000037:antagonists & inhibitors / Q000235:genetics*; D054334:Viral Regulatory and Accessory Proteins; D014779:Virus Replication
D006514:Hepatitis B Surface Antigens; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein