Synthesis of a mRNA vs Protein Vaccine: a Possible Treatment to Prevent Human Parainfluenza Virus in Children
Date
2019-12
Authors
Lohmann, Natalie
Major Professor
Advisor
Verhoeven, David
Committee Member
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Abstract
Human parainfluenza virus (HPIV) is the second most common viral respiratory pathogen in young children. It causes symptoms such as fever, cough, and runny rose and may lead to serious infections in the respiratory tract or secondary bacterial infections in the middle ears. Currently, there is no vaccine or antiviral treatment available. The goal of this project is to synthesize an mRNA vaccine to prevent HPIV, with initial focus on strain 3. mRNA vaccines represent a new way of vaccinating that are proving superior over DNA vaccinations and generally elicit better immune responses than protein vaccines. Here, viral RNA was extracted from HPIV type 3, reverse transcribed, and amplified for the two viral surface receptors HN and F proteins. cDNA was cloned into a T7 RNA polymerase expression system for manufacture of mRNA for vaccination. As a comparison against traditional protein vaccines, the same cDNA was also placed into a baculovirus expression system for manufacture of recombinant proteins for vaccination. After verification of expression of mRNA in mammalian cell lines and purification of viral proteins from insect cell lines, the potential vaccines will be utilized in mice with eventual testing of their antibodies against viral neutralization in cell culture.
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