Fig. 2
From: Multiplexed CRISPR/Cas9 gene knockout with simple crRNA:tracrRNA co-transfection
Multiplexed gene KO via crRNA combination. a Co-ablation of three target genes in U2OS-EGFP12-Cas9 cells. Cells were reverse transfected with the indicated crRNA:tracrRNA either individually or in combination. Protein expression was assessed by western at the indicated time points (Untreated, no transfection; NT, non-targeting control crRNA). b Quantification of target gene KO efficiency. Protein levels from a were quantified by densitometry. Parallel samples were collected at the indicated time points for target site indel analysis. The fraction of WT allele in the cell population was determined using TIDE (Combo, EGFP, PTEN, and KRAS crRNA combination). c The impact of CRISPR cutting on cell viability. Cell viability was determined 5 days after transfection using indicated crRNAs. (**p < 0.01 and *p < 0.05, two-tailed t-test)