Construction of an infectious clone for the chikungunya virus that lacks the furin cleavage site for the production of non-infectious particles

• Grant number: 18/11939-0
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: January 10, 2019
• End date: July 09, 2019
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Benedito Antônio Lopes da Fonseca
• Grantee: Danillo Lucas Alves Espósito
• Supervisor: Beate Kuemmerer
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: Universität Bonn, Germany
• Associated to the scholarship: 17/19137-7 - Study of Chikungunya virus immature particles - evaluation of infectivity, diagnostic approach and vaccine candidate, BP.PD

Abstract

The recent outbreak caused by chikungunya virus (Alphavirus, Togaviridae) worldwide has shown that, despite being first isolated in the 1950s, this infection still has several points to be elucidated. This especially includes the correct diagnosis of the disease caused by the virus, but also treatment and prevention of infection. The virus, which in spite of short-lived viremia, produces symptoms such as high fever, myalgia, rash, and arthritis/arthralgia, which may persist for months or years, causes a major public health impact. The present project proposes the construction of an infectious clone of chikungunya virus with a specific mutation that replaces the furin cleavage site in the structural protein region. The infectious clone will be produced after amplification and assembly of the complete viral genome, and the furin site will be replaced by site-directed mutagenesis. The resulting non-infectious particles will be analysed for viral attenuation by evaluating the infectivity in vitro in Vero and Hela cells to determine their potential use as vaccine candidate.