West Nile Virus (WNV) is a mosquito-borne virus that infects both animal and human populations. A genus of mosquitoes named Culex transmits WNV between different hosts with the primary vertebrate reservoir host being birds. The viral infection could be asymptomatic, cause West Nile fever, or cause a neuroinvasive disease such as meningitis, encephalitis, or acute flaccid paralysis. WNV entered the United States in 1999 taking form in an outbreak in New York City. There are currently no antiviral drugs or vaccinations against the virus, resulting in dramatically higher cases of WNV each year. Undeniably, WNV is a real threat to the entire world. The main goal of this project is to investigate the possibility of monitoring the activity of the viral protease in cells, which can be utilized for drug discovery. Once WNV, like other Flaviviridae viruses, causes infection and enters the cell, the virus encodes its genome as one single polyprotein. This polyprotein is cleaved by host and viral protease enzymes, which can then be transcribed and replicated. We thus hypothesize that inhibiting and blocking the cleavage process of the viral proteases can stop the WNV infection. The significance of our research is using a cell-based assay that tests and investigates the blockage and inhibition of protease activates in vivo, where all other previous studies were performed in vitro.