Investigating molecular mechanisms of dengue virus NS1 disruption of the glycocalyx-like layer and barrier dysfunction of human endothelial cells
Eva Harris, Professor
Public Health; Div of Infectious Diseases and Vaccinology
Closed. This professor is continuing with Fall 2024 apprentices on this project; no new apprentices needed for Spring 2025.
Previous studies in the Harris laboratory have demonstrated the ability of the secreted flaviviral protein, nonstructural protein 1 (NS1), to induce both hyperpermeability in vitro and vascular leak in vivo, both mediated by the disruption of endothelial glycocalyx components and intercellular junction degradation. Further work in the lab aims at the investigation of host factors involved in this process that might be attractive targets for therapeutic interventions. We employ a range of molecular and genetic methods like RNA-seq, immunoprecipitation-mass spectrometry and CRISPR/Cas9 to identify and modulate central pathways that might be involved in NS1 pathogenesis. Additionally, we are establishing systems to visualize host factors interacting with NS1 during its internalization and trafficking using high resolution microscopy. Current projects include the validation of target factors by interaction studies and the development of new tools to facilitate and optimize our imaging workflows. We are generating and characterizing knockout cell lines as well as cell lines expressing our proteins of interest and conducting mechanistic studies to elucidate the complete pathway of NS1-mediated endothelial barrier disruption. Further, we are producing recombinant NS1 with modifications of specific amino acids, domain substitutions, and containing a variety of tags to study protein-protein interactions.
Role: The URAP student will work under the direct supervision of graduate student Felix Pahmeier in Dr. Eva Harris’ laboratory. The student will be expected to work a minimum of 12-15 hours per week and commit to at least one year, ideally including the summer of 2022. This requires regular work on weekdays that may extend beyond after 5 pm and occasional work on weekends or as arranged with the postdoctoral researcher. Skills learned will include but not be limited to cell culture, immunofluorescence microscopy and molecular cloning. The URAP student is also expected to attend weekly research meetings and maintain detailed records of the work performed. The student is expected to complete the assignments in a timely manner, maintain open communication with other members of the research group and with the research coordinator, ask questions when in need of guidance, and actively ensure (through communicating with the research coordinator) that they are getting the experience they expect from the URAP program.
Qualifications: The apprentice must be at a sophomore or junior level and have a strong interest in molecular biology, biochemistry, and/or infectious disease. Experience with molecular biology and biochemistry is preferred. Other requisites are enthusiasm, high motivation, and the desire to develop independent thinking.
Day-to-day supervisor for this project: Felix Pahmeier, Staff Researcher
Hours: 12 or more hours
Related website: https://www.harrisresearchprogram.org
Biological & Health Sciences