RNA APTAMERS SPECIFIC FOR THE HCV NS3 PROTEASE AND HELICASE DOMAINS

Abstract

  The hepatitis C virus (HCV) non-structural protein 3 (NS3) is a multifunctional enzyme with protease and helicase activities. It is essential for HCV replication and proliferation and is therefore a target for anti-HCV drugs. To obtain efficient RNA aptamers specific for HCV NS3, and to develop inhibitors of HCV replication, we performed in vitro selection for the NS3 protease and helicase domains, respectively. In vitro selection, namely, SELEX (systematic evolution of ligands by exponential enrichment) is a useful strategy for isolating nucleic acid sequences that have a high affinity for a target molecule from a randomized oligonucleotide pool. Isolated RNA aptamers showed effective inhibition against either the protease or helicase activity of NS3 in vitro. In addition, new type of RNA ligands, which displayed dual-inhibitory functions targeting both NS3 protease and helicase activities were constructed. Furthermore, NS3 protease aptamers effectively inhibited NS3 protease activity in living cells.