Identification of Genes Regulating Gene Targeting by a High-Throughput Screening Approach

Figure 2

Representation of a typical primary screening run. Fourteen 96-well plates containing siRNAs for the screen and control siRNAs were transfected with RMLuc+I-SceI or pUC in duplicate. Seventy-two hours after transfection, luciferase activity was detected. Each dot represents the mean value per siRNA. The black box shows the luciferase value obtained following transfection with the empty vector (pUC), corresponding to background. Black triangles represent the values obtained for cotransfection with the AS siRNA and RMLuc+I-SceI; white circles represent the values obtained for cotransfection with RAD51 siRNA and RMLuc+I-SceI. Finally, white squares represent the values obtained for cotransfection with LIG4 siRNA and RMLuc+I-SceI. The black line indicates the mean value for the run. Dashed boxes show the cutoff values for hit definition.