No, while the transfection for siRNA and esiRNA are comparable, they are not identical. An siRNA transfection protocol may serve as a starting point for the separate esiRNA optimization process. Cell lines have different properties and are sensitive or insensitive to different manipulations. This is certainly also true for transfections. Therefore, for every cell line, an appropriate transfection protocol must be established separately. The goal of transfection optimization is to achieve maximal transfection efficiency with minimal cytotoxicity.
Provided below are the esiRNA transfection conditions for some commonly used cell lines. The values provided should only serve as guidelines (a detailed optimization should be carried out if other plate formats are used). We recommend using an esiRNA for Eg5 (KIF11) and Renilla luciferase (RLUC) as positive and negative controls, respectively, for optimization. Eg5 (KIF11) induces mitotic arrest with the formation of round-shaped cells that become apoptotic after several hours of arrest. These round-shaped cells can easily be observed in a standard bright-field cell-culture microscope. The conditions with a maximum number of round-shaped cells for the Eg5 transfection with a minimum cytotoxicity for the RLUC transfection signify optimal transfection conditions.
Figure 1: RLUC
Figure 2:mitotic arrest