Chikungunya Virus Glycoproteins Mediate Viral Entry and Cellular Fusion

The Chikungunya virus (CHIKV) has currently been identified in over 40 countries and in 2008 was listed as a US National Institute of Allergy and Infectious Diseases (NIAID) category C priority pathogen. Outbreaks of the virus have been documented as early as 1779 and frequent outbreaks have been reported through 1960-2003, most notably in Reunion Island, a French overseas department in the Indian Ocean. Out of a total population of 785,000 in Reunion Island, 300,000 cases were reported including a total of 237 deaths. Numerous aspects of the viral life cycle are unknown, with no current vaccine the implementation of more research and dissemination of more knowledge is of great public health importance. A CHIKV construct was synthesized by Genewiz, containing CHIKV structural proteins in pcDNA 3.1. This construct was used to create CHIKV pseudo-viral particles with a luciferase based reporter. The pseudo-virus was used to survey many cell lines for permissivity to infection. This construct was also used to create 3 other constructs containing CHIKV E1, E2, and E3 individually. These constructs were used individually and in combination with each other to create pseudo-viruses for cellular infection. The cell-cell fusion capabilities of the full CHIKV construct along with the individual envelope proteins were also tested in a Cre-Lox system