title: Evaluation of canonical siRNA and Dicer substrate RNA for inhibition of hepatitis C virus genome replication--a comparative study.
abstract: Hepatitis C virus (HCV) frequently establishes persistent infections in the liver, leading to the development of chronic hepatitis and, potentially, to liver cirrhosis and hepatocellular carcinoma at later stages. The objective of this study was to test the ability of five Dicer substrate siRNAs (DsiRNA) to inhibit HCV replication and to compare these molecules to canonical 21 nt siRNA. DsiRNA molecules were designed to target five distinct regions of the HCV genome - the 5' UTR and the coding regions for NS3, NS4B, NS5A or NS5B. These molecules were transfected into Huh7.5 cells that stably harboured an HCV subgenomic replicon expressing a firefly luciferase/neoR reporter (SGR-Feo-JFH-1) and were also tested on HCVcc-infected cells. All of the DsiRNAs inhibited HCV replication in both the subgenomic system and HCVcc-infected cells. When DsiRNAs were transfected prior to infection with HCVcc, the inhibition levels reached 99.5%. When directly compared, canonical siRNA and DsiRNA exhibited similar potency of virus inhibition. Furthermore, both types of molecules exhibited similar dynamics of inhibition and frequencies of resistant mutants after 21 days of treatment. Thus, DsiRNA molecules are as potent as 21 nt siRNAs for the inhibition of HCV replication and may provide future approaches for HCV therapy if the emergence of resistant mutants can be addressed.
authors.affiliation: Genomics Study Laboratory, Sao Paulo State University, IBILCE, São José do Rio Preto, SP, Brazil; School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, United Kingdom.
authors.forename: Bruno
authors.initials: B
authors.lastname: Carneiro
authors.affiliation: Genomics Study Laboratory, Sao Paulo State University, IBILCE, São José do Rio Preto, SP, Brazil.
authors.forename: Ana Cláudia Silva
authors.initials: AC
authors.lastname: Braga
authors.affiliation: Genomics Study Laboratory, Sao Paulo State University, IBILCE, São José do Rio Preto, SP, Brazil.
authors.forename: Mariana Nogueira
authors.initials: MN
authors.lastname: Batista
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, United Kingdom.
authors.forename: Mark
authors.initials: M
authors.lastname: Harris
authors.affiliation: Genomics Study Laboratory, Sao Paulo State University, IBILCE, São José do Rio Preto, SP, Brazil.
authors.forename: Paula
authors.initials: P
authors.lastname: Rahal
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D045744:Cell Line, Tumor; D016679:Genome, Viral / Q000235:genetics*; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D012093:Replicon / Q000235:genetics; D014162:Transfection / Q000379:methods; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014779:Virus Replication / Q000235:genetics*
chemical_list: D020121:5' Untranslated Regions; D034741:RNA, Small Interfering; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins