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

Figure 5

Characterization of EP300 and ATF7IP knockdown. Total RNA was extracted 48 hours after transfection with the indicated DNAs and siRNAs (two different siRNA sequences were used to target each of the ATF7IP and EP300 genes). RT-PCR was carried out to amplify mRNA transcribed from the ATF7IP, EP300, GAPDH, and I-SceI genes. The effects of siRNAs on their targets were monitored by analyzing band intensity after electrophoresis in 1% agarose gels. Lane 1, untransfected cells. In lanes 2 to 7, the cells were transfected with the plasmid carrying the repair matrix and the I-SceI expression cassette. Lane 2, no siRNA; lane 3, AS siRNA; lanes 4 and 5, siRNA against EP300; lanes 6 and 7, siRNA against ATF7IP.