Development of a pod pepper vein yellows virus-based expression vector for the production of heterologous protein or virus like particles in Nicotiana benthamiana

Plant viruses are emerging as a compelling alternative system for the heterologous production of pharmaceutical proteins, offering advantages in scalability, cost-effectiveness, and biological safety over traditional expression systems. They are increasingly recognized as effective platforms for biomedical applications, frequently used in the expression of human viral proteins and the display of peptides or proteins. The pod pepper vein yellows virus (PoPeVYV), classified within the genus Polerovirus of the Solemoviridae family, can substantially increase viral titers when co-infected with pod pepper vein yellows virus-associated RNA (PoPeVYVaRNA). This mixed infection methodology facilitates the formation of rod-shaped virus-like particles (VLPs), wherein modified green fluorescent protein (mGFP) is fused to the C-terminus of the coat protein (CP) from the pepper mild mottle virus (PMMoV). Notably, the expression of hepatitis B surface antigen (HBsAg) demonstrates a marked preference for plant viruses, allowing for enhanced expression via the PoPeVYV mixed infection system in Nicotiana benthamiana. Consequently, the PoPeVYV-based vector presents a promising alternative for the high-level production of heterologous proteins and rod-shaped VLPs in plants.