Inhibition of hepatitis B virus replication by shRNAs in stably HBV expressed HEPG2 2.2.15 cell lines.
In this study, the effect of RNAi on HBV replication was observed in a cell culture model, HepG2 2.2.15 cell line, which supports human HBV ayw replication and expression. Aim of the study was to investigate effects of shRNAs (small hairpin RNAs) targeting hepatitis B virus mRNAs on the viral replication in HepG2 2.2.15 cells. We selected three target HBV mRNA regions with different putative secondary structures to test whether the secondary structure of RNA may affect the inhibition efficacy on the target HBV RNA. Three HBV-specific siRNAs (small interfering RNA) were designed targeting X (1689-1708), Core (2229-2248) and S (765-784 nt) transcripts. HepG2 2.2.15 cells were transfected with shRNA expressing plasmids, P765, P2229 and P1689 targeting S, core and X region, respectively or a mock plasmid targeting lacZ gene. The culture media was collected throughout six days after transfection and analyzed by real-time PCR. Viral DNA production was suppressed for 7 days. The HBV DNA levels were decreased by 73, 72 and 79% with P765, P2229 and P1689 vectors, respectively. In conclusion, the shRNAs designed for X, core and S regions, specifically and significantly suppressed HBV DNA. siRNAs potentially may be used in treatment of hepatitis B.
Kayhan
H
H
Institute of Hepatology, Ankara University, Ankara, Turkey.
Karatayli
E
E
Turkyilmaz
A R
AR
Sahin
F
F
Yurdaydin
C
C
Bozdayi
A M
AM
D045744:Cell Line, Tumor; D018390:Gene Targeting; D015316:Genetic Therapy / Q000379:methods; D006515:Hepatitis B virus / Q000502:physiology*; D006801:Humans; D009690:Nucleic Acid Conformation; D034622:RNA Interference; D012333:RNA, Messenger; D034741:RNA, Small Interfering / Q000235:genetics*; D012367:RNA, Viral; D014162:Transfection; D014779:Virus Replication
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral