T cell transduction - (Nov/03/2009 )

I want to transduce human Tcells with a Foxp3 shRNA. I've heard that Tcells are very hard to transduce, does anyone have any tips?

Andrew on Nov 3 2009, 09:39 PM said:

I'd guess you could do it with a HIV-based lentiviral vector system, as it is already adapted to transducing non-dividing T-cells.

what vector backbone are you using? (lentivector or onco-retrovector)?

With activated T cells, I have gotten up to 70% transduction efficiencies with retrovirus infections. 60 minutes spinfection with 1 million cells and 1 mL viral sup. 8 ug/mL of polybrene.

I transduced 1-2 X 10^5 cells/well in 6-well plate with 1-2 ml lentivirus, spinoculation (900-1000 X g for 60min at room temp.) with 8 ug/ml polybrene, change medium after centrifugation.