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T cell transduction


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#1 Andrew

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Posted 03 November 2009 - 08:39 PM

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?

Edited by Andrew, 03 November 2009 - 08:40 PM.


#2 elpollodiablo

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Posted 04 November 2009 - 01:01 AM

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?


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.

#3 miBunny

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Posted 04 November 2009 - 06:41 PM

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.

#4 Functional Screens

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Posted 05 November 2009 - 06:17 PM

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.

Edited by Functional Screens, 05 November 2009 - 06:17 PM.





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