Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

Abstract

We have developed Cucumber mosaic virus (CMV) as a plant virus vector especially for production of pharmaceutical proteins. The CMV vector is a vector modifiable for different host plants and does not require further engineering steps. CMV contains three genomic RNA molecules (RNAs 1-3) necessary for infectivity. With this system, instead of creating different vector constructs for each plant we use, we take advantage of the formation of pseudrecombinants between two CMV isolates by simply reassembling a vector construct (RNA 2 base) and an RNA molecule containing the host determinant (mostly RNA 3). In this study, the gene for acidic fibroblast growth factor (aFGF), one of the human cytokines, was cloned under the control of the subgenomic promoter for RNA 4A of the CMV-based vector, C2-H1. Infected Nicotiana benthamiana plants produced aFGF at levels up to 5-8% of the total soluble protein. The tobacco-produced aFGF was purified, and its biological activity was confirmed. Using this system, which provides a versatile and viable strategy for production of therapeutic proteins in plants, we also demonstrated a high level of aFGF in Glycine max (soybean) and Arabidopsis thaliana.

Virus-Induced Necrosis Is a Consequence of Direct Protein-Protein Interaction between a Viral RNA-Silencing Suppressor and a Host Catalase

“…3). C2-H1 is a CMV-Y-based virus vector in which a multiple cloning site replaces the 2b gene (Matsuo et al, 2007). Therefore, the C-terminal region of HL2b is important for the interaction between 2b and CAT3.…”

Expression of The Human Acidic Fibroblast Growth Factor in Transgenic Tomato and Safety Assessment of Transgenic Lines

“…There are scientific reports about modifications made in the primary structure of aFGF for elimination of its strong mitogenic activity (70), for prolongation of its half-life and increased stability (73) which facilitate its use as a therapeutic protein (20,28,37). The gene for haFGF was transiently expressed in Nicotiana benthamiana (38,43), soybean and Arabidopsis (43).…”

Virus-induced gene silencing (VIGS) in <i>Lilium leichtlinii</i> using the <i>Cucumber mosaic virus</i> vector

“…A typical feature of CMV is its wide host range that includes the Liliaceae (Masuta et al 2002), and the Lilium-derived CMV-HL strain (Hagita et al 1989;Masuta et al 2002) is expected to be able to induce gene silencing in lilies. CMV is well characterized as a VIGS vector in other plant species (Hong et al 2012;Kanazawa et al 2011;Liu et al 2010;Matsuo et al 2007;Nagamatsu et al 2007;Otagaki et al 2006). CMV possesses tripartite positive-sense RNA genomes (RNA1, 2, and 3) and the 2b protein in the RNA2 genome acts as a silencing suppressor (Nakahara and Masuta 2014).…”