Engineering of an Aptamer Tagged Dengue Virus for RNA Imaging
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Author:
Lombard, Clair
Area of Honors:
Biochemistry and Molecular Biology
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
Joyce Jose, Thesis Supervisor Timothy Charles Meredith, Thesis Honors Advisor
Keywords:
Dengue Virus Flavivirus Life Cycle RNA Imaging Techniques
Abstract:
Dengue Virus, a member of the Flavivirus genus, is transmitted by Aedes mosquitoes and accounts for roughly 400 million infections and 40 000 fatalities annually. As there are currently no effective antivirals or vaccines available, the control of DENV has been largely vector focused. This thesis discussed the engineering of an aptamer tagged Dengue virus for RNA imaging which can aid in the understanding of viral mechanisms. Due to the complexity of flavivirus assembly mechanisms, traditional imaging methods have fallen short in providing the necessary molecular insights. This work uses the fluorogenic RNA aptamer Mango II, which emits a fluorescent signal upon binding to tagged RNA in the presence of TO1-biotin dye. The high-resolution capabilities of Mango II, along with its compatibility with live cell imaging and Streptavidin pull-down assays, offer an approach to studying RNA dynamics in the context of Dengue virus. The development of such a clone will be the first time that the Dengue virus RNA could be fully visualized, and findings from such a clone could provide a pivotal step in understanding the RNA-protein interactions during the flavivirus life cycle and potentially aid in the development of antiviral strategies.
