Study Of Novel Methods To Inhibit The Reproduction And Translation Of Hepatitis C Virus

Hepatitis C virus (HCV), a positive single-stranded RNA virus, is a major cause of liver disease in humans. In this paper, we report a novel strategy to inhibit the reproduction and translation of HCV with a nucleic acid electrochemical biosensor. Based on this biosensor, we also develop a method to quantitatively detect NF-кB. This method may be a good candidate for assaying NF-кB in the future.1. Study of novel methods to inhibit the reproduction and translation of Hepatitis C virusWe report a novel strategy to inhibit the reproduction and translation of HCV by using a piece of RNA, named as an additional RNA, which may activate the endonuclease activity of Argonaute2(Ago2) protein. With the help of the additional RNA, HCV genome RNA and the additional RNA may have12Watson-Crick bases pairing with microRNA-122(miR-122), which then activates the endonuclease activity of Ago2protein, resulting in the cleavage and release of HCV genome RNA from Ago2and miR-122. Consequently, the reproduction and translation of HCV is efficiently inhibited without the involvement from Ago2and miR-122. To assay the cleavage of the HCV genome RNA by Ago2, we designed an electrochemical biosensor with S1nuclease, which hydrolyzes single stranded regions in DNA-RNA hybrids. With this electrochemical biosensor, we can reveal that Ago2protein was activated only in the presence of additional RNA. So, this study may present an alternative method to inhibit HCV, and hold great promise in HCV treatment in the future.2. A nucleic acid electrochemical biosensor to detect NF-кB based on exonucleaseBased on the nucleic acid electrochemical biosensor fabricated in the second section, we modified the complex of G-quadruplex and NF-кB specific binding sequences on the electrode surface. With the help of λ exonuclease, G-quadruplex was formed in the presence of hemin. With the electrochemical signals of hemin, we can quantitatively detect NF-кB. The proposed method might become a promising approach for NF-кB assay.