Identifying Initiation Sites of Innate Immune Signaling during Paramyxovirus Infection

David Yang working at microscope

Students

2020
College

Project Summary

The Paramyxoviridae is a family of viruses that cause various important respiratory illnesses in humans and animals including diseases such as mumps, human parainfluenza virus, and respiratory syncytial virus (RSV). As they replicate in the host cell, many of the Paramyxoviruses produce defective viral genomes (DVGs), truncated versions of the full viral genome that while unable to replicate on their own, stimulate a strong immune response. The goal of my project is to determine if the cellular distribution of defective and full length viral genomes contributes to their respective abilities to induce an immune response in the host cell. To accomplish this goal, I am using a technology called bimolecular fluorescence complementation (BiFC) which will allow me to visualize where signal transduction of a set of anti-viral innate immune proteins called the RIG-I like receptors (RLRs) is occurring. I will then use imaging for viral and defective genomes to determine their locations relative to the RLR signaling. I am currently working on constructing this BiFC system through molecular cloning which will eventually be inserted in cells and coupled with viral infection. Ultimately by using imaging for the RLR protein signaling and viral genomes, we should be able to see exactly where RLR signaling occurs in a cell during the viral infection and see based on locations of signaling and viral genomes whether defective or full-length genomes play a larger role in initiating the immune response. By working on this project, I have learned to perform and understand the science behind various techniques commonly used in molecular biology and virology, such as PCR, RNA extraction, quantitative PCR, gel electrophoresis, DNA plasmid transfection, viral infection. Additionally, I have also gained critical thinking skills by working on experimental design and optimizing experiments. Overall, working in a research lab has really allowed me to explore and expand my passions for virology and immunology beyond simply learning material in a classroom or textbook.