1. Academic Validation
  2. Generation of Combinatorial Lentiviral Vectors Expressing Multiple Anti-Hepatitis C Virus shRNAs and Their Validation on a Novel HCV Replicon Double Reporter Cell Line

Generation of Combinatorial Lentiviral Vectors Expressing Multiple Anti-Hepatitis C Virus shRNAs and Their Validation on a Novel HCV Replicon Double Reporter Cell Line

  • Viruses. 2020 Sep 18;12(9):1044. doi: 10.3390/v12091044.
Hossein M Elbadawy 1 Mohi I Mohammed Abdul 1 Naif Aljuhani 1 Adriana Vitiello 2 Francesco Ciccarese 2 3 Mohamed A Shaker 4 5 Heba M Eltahir 1 Giorgio Palù 2 Veronica Di Antonio 2 3 Hanieh Ghassabian 2 Claudia Del Vecchio 2 Cristiano Salata 2 Elisa Franchin 2 Eleonora Ponterio 2 6 Saleh Bahashwan 1 Khaled Thabet 7 Mekky M Abouzied 1 7 Ahmed M Shehata 1 8 Cristina Parolin 2 Arianna Calistri 2 Gualtiero Alvisi 2
Affiliations

Affiliations

  • 1 Department of Pharmacology and Toxicology, College of Pharmacy, Taibah University, Almadinah Almunawwarah 41477, Saudi Arabia.
  • 2 Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.
  • 3 Immunology and Molecular Oncology Unit, Veneto Institute of Oncology IOV-IRCCS, 35128 Padua, Italy.
  • 4 Pharmaceutics and Pharmaceutical Technology Department, College of Pharmacy, Taibah University, Almadinah Almunawwarah 41477, Saudi Arabia.
  • 5 Pharmaceutics Department, Faculty of Pharmacy, Helwan University, Cairo 11795, Egypt.
  • 6 Fondazione Policlinico Universitario "A. Gemelli"-I.R.C.C.S., 00168 Rome, Italy.
  • 7 Department of Biochemistry, Faculty of Pharmacy, Minia University, Minia 61519, Egypt.
  • 8 Department of Pharmacology and toxicology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62511, Egypt.
Abstract

Despite the introduction of directly acting antivirals (DAAs), for the treatment of hepatitis C virus (HCV) Infection, their cost, patient compliance, and viral resistance are still important issues to be considered. Here, we describe the generation of a novel JFH1-based HCV subgenomic replicon double reporter cell line suitable for testing different Antiviral drugs and therapeutic interventions. This cells line allowed a rapid and accurate quantification of cell growth/viability and HCV RNA replication, thus discriminating specific from unspecific Antiviral effects caused by DAAs or cytotoxic compounds, respectively. By correlating cell number and virus replication, we could confirm the inhibitory effect on the latter of cell over confluency and characterize an array of lentiviral vectors expressing single, double, or triple cassettes containing different combinations of short hairpin (sh)RNAs, targeting both highly conserved viral genome sequences and cellular factors crucial for HCV replication. While all vectors were effective in reducing HCV replication, the ones targeting viral sequences displayed a stronger Antiviral effect, without significant cytopathic effects. Such combinatorial platforms as well as the developed double reporter cell line might find application both in setting-up anti-HCV gene therapy approaches and in studies aimed at further dissecting the viral biology/pathogenesis of Infection.

Keywords

antivirals; gene therapy; hepatitis C virus; reporter cell line; siRNA.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-15005
    99.97%, HCV 抑制剂
    HCV