Retroviral-Mediated Transduction: A Method to Introduce Target Gene into Cancer Cells

- Retroviruses are RNA viruses that replicate by using reverse transcriptase to convert their RNA genome to double-stranded DNA and permanently integrate into the host genome. New viral particles exit the cell and can infect other cells. Cancer gene therapy employs retroviral vectors to introduce therapeutic genes into target cells via a technique called transduction to downregulate the mutated gene or eliminate cells.

To obtain high-titer retroviral vectors, transfect the HEK 293 cells with a transfection mixture containing the desired retroviral plasmid, packaging plasmid, and a suitable transfecting agent. Incubate the cells. The transfecting agent facilitates the delivery of both plasmids into the cells.

The retroviral plasmid encodes the viral genome with the target gene while the packaging plasmid encodes the viral envelope proteins. The packaging cells' cellular machinery forms retroviral particles and releases them into the medium. Collect the viral supernatant and centrifuge to remove cells.

Transfer the desired concentration of viral particles and cancer cells to a fibronectin-coated culture plate to facilitate binding viral envelope proteins to cells. Incubate the cells.