Figure 1. | Schematic showing production of lentiviral particles using the Trans-Lentiviral Packaging System
Figure 2. | Lentiviral vector particles were produced from three different GIPZ clones using both the Arrest-In and Calcium Phosphate (CaPO4) transfection reagents. To generate the viral particles, HEK293T cells were co-transfected with the individual GIPZ constructs and the Trans-Lentiviral Vector packaging mix following recommended protocols. Culture supernatants containing the viral particles were collected approximately 64 hours post-transfection.
Supernatants were clarified by low-speed centrifugation, aliquoted, and stored at -80°C. To determine functional titers, HEK293T cells were seeded in a 24-well plate and transduced the following day with a dilution series of each of the viral particles. At 72 hours post-transduction, the transduced cells (TurboGFP-positive) were counted under fluorescence microscopy and titers calculated as transducing units per milliliter (TU/ml). Results indicate that transfection using CaPO4 produced higher titers with all three GIPZ clones as compared to Arrest-In transfection.