Figure 3

Target site and knockout efficiency of gRNAs. (a) Schematic presentation of gRNA target location on the Venus transgene. Nine gRNAs were designed to target the promoter region (− 252, − 72, and − 69), 5′ region (36, 100, and 121), and 3′ region (518, 554, and 676) of Venus transgene which are depicted with white-green, black, and yellow bars, respectively. The target site of the digital PCR assay is depicted in a red box. (b) Efficiency of different gRNAs for making Venus knockout. Cells were treated with puromycin and were screened for the Venus transgene under a fluorescence microscope 10 days after the electroporation (n > 3). Bars with common letters are not significantly different (p-value < 0.05). Cells with one copy of Venus, which did not undergo any electroporation treatment, were considered as the positive control for the Venus expression (PK).