An optimized conditional suicide switch using doxycycline-dependent expression of human tBid.
We have examined in a HeLa model system whether tight doxycycline-dependent expression of human tBid can be used as a fast and efficient suicide switch. A stably transfected cell line expressing human tBid in a strictly dox-dependent manner showed fast and efficient killing with up to 98 % specific cell death 24 h after induction. The survival rate was only 0.6 % after eleven days of dox treatment. Very low amounts of tBid were sufficient to induce apoptosis indicating that stringent control of the dox-dependent suicide gene is essential for cell survival in the absence of inducer. Using human tBid, an endogenous effector protein with low immunogenic potential, and doxycycline, an inducer with low cytotoxicity and a long record of safe use in humans, this conditional suicide switch provides a tool for various applications, e.g., adoptive immune transfer.
Knott
Anja
A
Lehrstuhl für Mikrobiologie, Friedrich-Alexander-Universität, Erlangen-Nürnberg Staudtstr. 5, Erlangen, Germany.
Drueppel
Lars
L
Beyer
Thomas
T
Garke
Kristin
K
Berens
Christian
C
Herrmann
Martin
M
Hillen
Wolfgang
W
D017209:Apoptosis / Q000187:drug effects*; D051023:BH3 Interacting Domain Death Agonist Protein / Q000378:metabolism*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004318:Doxycycline / Q000494:pharmacology*; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D010957:Plasmids; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D014162:Transfection
D051023:BH3 Interacting Domain Death Agonist Protein; D049452:Green Fluorescent Proteins; D011065:Poly(ADP-ribose) Polymerases; D004318:Doxycycline