Artificial Zinc Finger Protein Designed To Target X Gene Promoter Of HBV And Inhibits The Transcription Of Hbv In HepG2.2.15Cells

Objective： To observe the inhibition of hepatitis B virus(HBV)transcription in vitro by zinc finger protein(ZFP)which was designedspecifically to target X gene promoter of HBV(HBX).Methods：Recombinant plasmid pcDNA3.1-ZFP and pcDNA3.1-NCwere transfected into HepG2.2.15cell by LipofectamineTM2000as ZFPgroup and NC group.1. The expression of HBX protein was detected by Western blot24hours after transfected.2. Hepatitis B e antigen (HBeAg) levels in supernatant ofHepG2.2.15cells24hours after transfected with pcDNA3.1-ZFP weredetected by ELISA.3. HBV DNA levels in supernatant of HepG2.2.15cells aftertransfected with pcDNA3.1-ZFP were detected by q-PCR.4. HBV mRNA was tested by PT-PCR.Results：Recombinant plasmid pcDNA3.1-ZFP was transfected intoHepG2.2.15cells.1. Compared to the control group, the expression of HBX protein ofthe ZFP group was reduced.2. In the presence of ZFP, the level of HBeAg was reduced by33.8%. 3. In the presence of ZFP, HBV DNA level was considerably reducedby51.7%4. Compared to the control group, HBV mRNA was sharply decreased.Conclusion: These results show that artificial zinc finger proteindesigned to target X gene promoter of HBV can inhibit the transcription ofHBV in HepG2.2.15cells effectively.