...described above (envelope, packaging, and transfer) are co-transfected into the HEK293T packaging cell line...plasmids containing the transfer, envelope, or packaging components. The transfer plasmid encodes the gene...gamma-retroviral vectors, you need three plasmids (Figure 2): Transfer plasmid — contains transgene, sgRNA, or shRNA ...transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most standard cloning...restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited restriction sites or... number of plasmids that are required in the transfection step of the viral packaging cell line. For example...events occur between the packaging, envelope, and transfer plasmids, and that resulting construct is packaged...