Uptake of Modified Red Clover Necrotic Mosaic Virus and Small Molecule Release from the Virion

This research seeks to investigate the interactions between human cervical cancer cells (HeLa) and a novel synthetic drug delivery vector. This delivery vector is constructed by labeling a plant virion—Red Clover Necrotic Mosaic Virus (RCNMV)—with multiple copies of a fouorescein-labeled adenoviral nuclear localization signal (NLS) peptide. Rhodamine dye is infused into the labeled virion, and the construct is delivered to HeLa cells so that cellular uptake and rhodamine release inside the cell can be simultaneously monitored using fluorescence activated cell sorting (FACS) flow cytometry. It has been found that cellular uptake of the modified RCNMV constructs is a function of incubation time and rhodamine concentration inside the virion. The maximum uptake is seen after a six hour incubation time with virions containing an average of five rhodamine molecules. Furthermore, it has been determined that the amount of rhodamine released from the virion increases with incubation time. Again, the maximum amount of rhodamine release is seen after a six hour incubation time. Once this system is fully characterized, animal studies of virion localization, small molecule drug encapsulation in the virion, and deliveries to more selective cell lines to demonstrate potential cell specificity of the construct will be performed.