Research Symposium

BIO

I am a IMS Clinical Professions major on a pre medical route. I grew up in Cocoa Beach, Fl where I received my Associates Degree while still in high school. This process allowed me to explore my interests within the medical field, specifically orthopedic surgery and oncology. Research has also been an interest of mine, and having the opportunity to work in the Francis Lab has allowed me to explore the ideals of working in a lab and creating on my own projects. This is the first research lab I have worked in at Florida State and I plan on continuing in this lab for the rest of my time at FSU.

Construction and Validation of a Halo-tagged fluorescence HIV-1 particles for single virus punctate imaging inside living cells

Abstract

A Halo-tag is a self-labeling protein tag used to study interactions and localization through fluorescence imaging. My UROP project was geared towards creating a Halo-tagged virus that would have the unique capabilities of fluorescing at two different wavelengths. Once the tagged virus is mixed with the cells, a microscope will enable the visualization of where the virus is and how it travels around the cell. To accomplish this goal I constructed a plasmid encoding for the Vpr-integrase-Halo tag (BruVIN-Halo) fusion protein, that when co-transfected with a viral genome encoding plasmids can become incorporated into virions. I evaluated the incorporation of the BruVIN-Halo, and its labeling by Janelia Fluorophore (JF646) by confocal microscopy. Image analysis showed a strong labeling of the viral BruVIN-Halo protein, which appeared as a bright puncta resistant to photobleaching over ~4000 iterations of imaging. In ongoing work, I am evaluating the effects of bruVIN-Halo tag incorporation on virus infection. Subsequently, I am to infect living Hela Cells and image the virus localization and trafficking in the nucleus. These experiments will help me accurately pinpoint the localization of HIV-1 inside different compartments of the mammalian cell. I also aim to explore stochastic labeling of the IN-Halo protein, which involves colabelling the virus with 2 fluorophores, and examine molecular changes to the virus architecture by fluorescence resonance energy transfer (FRET) imaging of HIV-1 infection.

Keywords: Virology, Halo-tag, fluorescence, HIV-1