Mutant Nipah Virus-like Particles as a Protein Delivery Technology
Open Access
- Author:
- May, Luke
- Area of Honors:
- Immunology and Infectious Disease
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Anthony Paul Schmitt, Thesis Supervisor
Robert Paulson, Thesis Honors Advisor - Keywords:
- Virus-like particles
Nipah virus
protein delivery technologies - Abstract:
- Gene and protein delivery technologies are a critical therapeutic in the treatment of genetic diseases. Proteins like CRISPR-Cas9 could eventually be delivered to edit genes and treat a wide variety of disorders. Virus-like particles (VLPs) are one promising method to deliver proteins. These non-infectious particles are composed of viral components and are capable of enclosing and transporting a variety of functional proteins. Paramyxoviruses, a class of enveloped RNA viruses, have been used for VLP research. Interactions between the matrix (M) and nucleocapsid (NP) proteins drive the packaging of cargoes and budding of particles. Using parainfluenza virus 5 (PIV5), previous research has shown that a 30 amino acid sequence from the C-terminal end of PIV5 NP protein can confer packaging efficiency to a protein of interest when bound to it. PIV5 M protein amino acid substitutions made to areas critical for M-NP interactions resulted in increased VLP production. However, a PIV5-based VLP therapeutic may meet challenges in practice due to preexisting immunity in a significant proportion of the global population. Nipah virus (NiV), a zoonotic paramyxovirus that is genetically homologous to PIV5, can also create VLPs and has less risk of preexisting immunity. In this study, Nipah virus M protein mutants analogous to those that improved PIV5 VLP formation were generated and tested for increased VLP production compared to wild-type Nipah virus M protein. From a group of 22 proposed mutants, only one, Y187L, showed improvement in VLP production compared to wild type. These results are important for future efforts to maximize the yield of VLPs in mammalian cell lines to produce therapeutics.