TransIT®-Lenti transfection reagent is designed to enhance delivery of packaging and transfer vectors to adherent HEK 293 cell types.

• Increase recombinant lentivirus production
• Greater than two fold higher functional titers are achieved over competing transfection reagents
• Enhance delivery of vectors to adherent cell types

TransIT®-Lenti Transfection Reagent is designed to enhance delivery of packaging and transfer vectors to adherent HEK 293 cell types to increase recombinant lentivirus production

To produce lentivirus, cells are transfected with packaging plasmids encoding the required gag, pol, and rev structural and regulatory genes as well as a transfer vector encoding the gene of interest (GOI). Replication incompetence is achieved by expressing the minimal virus components on separate plasmids and incorporating the self in-activating (SIN) elements through a major deletion of the 3’ long terminal repeat (LTR) within the transfer vector. Essential components are combined with the VSV envelope protein G for a broad virus tropism and increased stability during purification procedures. The lentivirus particles are secreted into the culture medium where they are collected, filtered and frozen into aliquots for subsequent transduction into target cells.

TransduceIT™ Reagent is an aqueous solution of hexadimethrine bromide, a cationic polymer, that is shown to greatly enhance retroviral transduction and transgene expression in both dividing and quiescent mammalian cells. Final TransduceIT™ Reagent concentrations of 5—10 μg/ml are appropriate for most transductions, although the optimal concentration for each cell type should be empirically determined.

HEK 293T cells are commonly used for lentivirus production due to high transfection efficiencies and protein expression levels; however, the lentivirus platform is often limited by insufficient viral titers requiring concentration before use. With TransIT®-Lenti, greater than twofold higher functional titers are achieved over competing transfection reagents.