siRNA transfection - (Jul/01/2005 )
I will transiently transfect my cell with siRNA oligo. I plan to dected the expression of target gene and its protein 1-5 days post transfection. Because cell will devide, will this lead to the more cells do not contain siRNA? If so, do I need transfect siRNA everyday during 5-day analysis.
How long the effect of a transiently transfected siRNA will last depends on the protein you want to "knock", the efficiancy of the siRNA, your cells, the transfection efficiancy,.... .
First thing I would do is to transiently transfect your cells and take samples every 12-24 hours and analyze by western-blot. That gives a clue how long your siRNA will work in your cells with your transfection efficiancy................ .
I would try to avoid to knock your cells everyday!
This might annoy your cells as most of the transfection procedures mean a kind of stress to them and may give you results in the further analysis which are not due to your siRNA´s but a consequence from the stress you gave your cells.