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Description
Human Immunodeficiency Virus (HIV), the causative agent of AIDS, has resulted in more than 33 million deaths since its discovery. Affecting people of various ethnic backgrounds, gender, and economic class, HIV is a prime example of a global health threat. With an alarming 50,000-estimated new cases per year there remains a clear need for continued HIV based-research as well as the development of novel therapeutics for infected individuals, especially those in poorer countries with limited access to current medicine. Furthermore, discovering and establishing new targets for antiviral development will help researchers and medical professionals alike in the fight against HIV/AIDS. Throughout the life cycle of most viruses there are instances of direct interaction between the host cell and the virus. These "cross-talk" events are often times ideal candidates for novel drug development efforts. The secretory pathway in eukaryotic cells is a great example of a site where direct viral-host interactions occur. Many RNA viruses utilize the vesicles of the secretory pathway as the site for maturation of their viral glycoproteins while they are being transported to the cell surface. HIV is a great example, as it uses the secretory pathway for processing and transport of its envelope polyprotein, which is required for HIV to produce infectious viral progeny and continue the spread of the virus. Therefore, blocking or disrupting this process of HIV Envelope (Env) maturation represents an attractive target for drug development efforts. Historically, cell-based platforms have served as valuable tools for targeted or phenotypic-based drug discovery. Cell-based assays offer researchers a wide-range of benefits during the early stages of drug development such as identifying cytotoxic chemicals and membrane-traversing compounds. These advantages can result in saving valuable time as well as money for scientists. The work outlined here describes the adaptation of an established cell-based assay, engineered to monitor cleavage of HIV Env within the vesicles of the secretory pathway, to larger segments of HIV Env. These larger peptides will facilitate the search for compounds that disrupt Env processing and/or transport to the cell surface