Intracellular Delivery of HCV NS3p gene using vectored particles

Author(s)

Nobre Lemos, Marcos Alexandre ^[1] ; Suarez Patino, Sandra Fernanda ^[1] ; Bernardino, Thaissa Consoni ^[1] ; Coroadinha, Ana Sofia ^[2] ; Soares, Hugo ^[2] ; Astray, Renato Mancini ^[1] ; Pereira, Carlos Augusto ^[1] ; Calil Jorge, Soraia Attie ^[1]

Affiliation

^[1] Inst Butantan, Lab Imunol Viral, Ave Vital Brasil 1500, BR-05503900 Sao Paulo - Brazil

^[2] iBET, Cell Line Dev & Mol Biotechnol Lab, Ave Republ, Oeiras - Portugal

• Document type: Journal article
• Source: Journal of Biotechnology; v. 274, p. 33-39, MAY 20 2018.
• Web of Science Citations: 0

Abstract

Viral hepatitis caused by the hepatitis C virus (HCV) affects millions of people worldwide. The non-structural protein 3 (NS3), one of the most conserved proteins in HCV, is the target of many therapeutic studies. The NS3 protease domain (NS3p) has a range of cytotoxic T lymphocyte (CTL) epitopes, and synthesizing the protein inside the cells is the most appropriate way to present it to the immune system. We developed a tool to study this kind of presentation, using two vectored particle (VP) systems, one based on the Semliki Forest virus (SFV) and the other on HCV pseudoparticles (HCVpp), both carrying the protease domain of the NS3 gene. In addition to producing the particles, we developed a method to quantify these VPs using qRT-PCR. We produced batches of approximately 2.4x10(4) SFV-NS3p/mu L and 4.0x10(2) HCVpp-NS3p/mu L. BHK-21 and HuH-7 cells treated with the VPs expressed the NS3 protein, thus showing the functionality of this system. (AU)

• FAPESP's process: 09/08038-1 - Gene Expression using viral pseudparticles
• Grantee: Soraia Attie Calil Jorge
• Support Opportunities: Regular Research Grants