INTRACELLULAR INHIBITION OF VIRAL GENE EXPRESSION BY HAMMERHEAD RIBOZYMES AGAINST HEPATITIS C VIRUS 5′ NCR AND C REGIONS / 解放军医学杂志

ABSTRACT

To investigate the potential use of synthetic stabilized ribozymes for the treatment of chronic hepatitis C virus (HCV) infection,we designed and synthesized 2 hammerhead ribozymes (Rz213 and Rz498) targeting conserved sites in the 5′noncoding region (NCR) and C gene of HCV RNA.Constructed to the eukaryotic vector pcDNA3, the two ribozymes were respectively or simultaneously transfected with lipofectamine into WISHnc transgenic cells, which could express permanently HCV C luciferase protein under the control of HCV 5′NCR.The expression of C luciferase was measured by luminometer.The results showed that the luciferase activities were significantly down regulated in the WISHnc cells, and the inhibitory rates were 42.94%～67.81% within 7 days after ribozymes transfection. There was no significant differences between Rz213 or Rz498 and co transfection, but adding the target site of ribozymes might prevent host cells from the loss of ribozyme therapeutic effect due to viral gene mutation.