Expression and characterization of plant produced AHSV nanoparticles encapsulating EGFP

Martins, Angelo

Expression and characterization of plant produced AHSV nanoparticles encapsulating EGFP

Thesis / Dissertation

2024

Abstract

Virus-like particles (VLPs) are virus-based nanoparticles that resemble the native virion but do not contain the viral genome. Heterologous expression of the viral structural proteins results in the spontaneous self-assembly of VLPs that have an empty inner cavity. These particles have an organized, repetitive structure that is very amenable to modification. One can take advantage of this property and fuse a foreign molecule to one of the viral structural proteins and upon VLP self-assembly this foreign molecule may be encapsulated within the empty inner cavity of the VLP. One can also take advantage of the high tailorability of VLPs and functionalize the external surface with targeting ligands for delivery purposes. Alternatively, the structural proteins that form VLPs can have inherent amino acid motifs with a natural targeting affinity for receptors that may be overexpressed on the surface of certain types of cells, for example cancer cells. These properties impart VLPs with great potential as delivery vehicles of diagnostic and/or therapeutic molecules. Core-like particle (CLPs) are a derivative of VLPs that share all the same properties and modification potential, however unlike VLPs, CLPs are comprised of only the viral structural proteins that form the inner capsid layer or ‘core' of the native virion. Herein, two cloning techniques have been used to successfully fuse a fluorescent protein, enhanced green fluorescent protein (EGFP), to the VP3 structural protein of African horse sickness virus (AHSV). Plant-based transient expression of the AHSV EGFP-VP3 and AHSV VP7 structural proteins has been used to generate CLPs from N. benthamiana leaves that successfully encapsulate the EGFP protein. However, interaction of this particle with the human integrin receptor, αvβ3, a receptor that is commonly found overexpressed on the surface of cancer cells could not be confirmed. These results highlight the potential of AHSV CLPs as a cargo carrier, but more research is required to elucidate its potential as a delivery vehicle

Keywords

Molecular and Cell Biology

Reference:

Collections

Masters

Full item page