chloroquine to improve transfection - useful or superfluous? (Apr/13/2007 )
in a protocol for Ca2+-precipitation/transfection it is recommended to add 2h after transfection some chloroquine solution to inhibit lysosomal DNA; I think it should improve transfection; has any one experience with and w/o chloroquine? does it really help to improve transfection? isn´t it cytotoxic?
The toxicity for CQ is cell type-dependent. I have not used myself in conjunction with Ca/Pi. You may also try glycerol or DMSO osmotic shock. I would say that the overall injury effect will add up for each component and treatment.
CQ does little for most lipid, or PEI-based transfection reagents.
I usually do not use chloroquine in CaPO4 transfections, only with DEAE-dextran. It does make a difference but can be toxic. Don't leave it on your cells for more than 3 hours, so for CaPO4, add it 3 hours before you plan to harvest.. I also do a 2 min, 10% DMSO (in PBS) "shock" after the transfection and from my experience of accidently forgetting to do that step, it makes a BIG difference.
thanks for nice suggestions but what is the exact time point for the DMSO shock? do you mean with "after transfection" after incubation with CaPO4?
Yes, I do the DMSO shock after the 6 hour CaPO4 incubation.