Lentivirus Production Protocol v1

This protocol describes producing lentiviral particles by transiently transfecting HEK-293 based packaging cells with a transfer vector plus packaging and envelope plasmids, harvesting and clarifying viral supernatant, and then transducing target cells in a 6-well format. Transduced cells are recovered in fresh media and subsequently selected to generate a stable, enriched population before expansion outside the lentivirus workspace. Expected results include robust viral production from high-transfection-efficiency HEK-293T or HEK-293FT cells and efficient gene delivery to target cells, enhanced by polybrene during transduction. After antibiotic selection, surviving cells should show stable integration and consistent expression of the lentiviral construct (e.g., fluorescent reporters or other transgenes) with minimal residual non-transduced cells.