Lentiviral Vector Production Protocol

Lentiviral vector production was performed as previously described 51 (link). In brief, HEK293T cells at a confluency of 70-90% were transfected with DNA:MACSfectin™ (Miltenyi Biotec) complexes using a total amount of 50µ DNA per T175 flask. HEK293T cells were then incubated overnight and sodium butyrate was added at a final concentration of 10 mM. Lentiviral vector containing supernatant was collected 48-60 h after transfection and following a filtration through 0.45 μm-pore-size PVDF filters subjected to centrifugation at 4 °C and 4,000×g for 24 h. Air-dried pellets containing lentiviral particles were resuspended at a 200-fold concentration with 4 °C cold PBS, aliquoted and stored at -80 °C.