Figure 3

From: siRNA silencing of keratinocyte-specific GFP expression in a transgenic mouse skin model

Figure 3

Identification of potent and selective siRNAs targeting reporter expression. (a) siRNA screening. Candidate siRNAs (CBL1-10) were identified using the Dharmacon siRNA prediction algorithm and evaluated in human 293T cells by co-transfection with a phMGFP/CBL plasmid that expresses hMGFP and CBL from a bicistronic mRNA (CMV promoter). The effectiveness of each analyzed siRNA (1 nM) was evaluated 48 h after co-transfection by quantitating bioluminsescence after luciferin substrate addition using the IVIS 50 Imaging system, as described in Materials and methods. An irrelevant siRNA (targets unrelated EGFP with no sequence similarity to hMGFP) was used as a negative control. (b) Dose response of CBL3 siRNA. CBL3 siRNA, at the indicated concentrations, was cotransfected with phMGFP/CBL plasmid in 293T cells and its effectiveness determined, as described above. Red depicts highest luciferase expression levels, blue the lowest.