Figure 2
From: A surrogate reporter system for multiplexable evaluation of CRISPR/Cas9 in targeted mutagenesis
Assessment of the sgRNA efficiency by the UDAR assay in HeLa and HEK293T cells. (a) Representative flow cytometry plots of HeLa cells co-transfected with a universal donor and sgRNA. (b) Representative results of the T7E1 assay (three replicates are presented for each assay) at the CSPG4 locus in the HeLa cells. Uncut (709 bp) and cut (537 bp and 172 bp) PCR bands are indicated. The indel ratios were calculated according to the band intensities. (c) Assessment of an sgRNA targeting the CSPG4 gene by the UDAR and T7E1 assay in HeLa cells. EGFP percentages analysed by FACS (green) and indel ratios, measured by the T7E1 assay (grey), are plotted in the same graph. The error bars indicate s.d. (n = 3). (d) Correlation between EGFP percentages and indel ratios at the CSPG4 locus in HeLa cells. Pearson’s correlation coefficient (R) = 0.81. (e) Assessment of an sgRNA targeting the LRP1 gene by the UDAR and T7E1 assay in HEK293T cells. EGFP percentages analysed by FACS (green) and indel ratios, measured by the T7E1 assay (grey), are plotted in the same graph. The error bars indicate s.d. (n = 3). (f) Correlation between the EGFP percentages and indel ratios at the LRP1 locus in the HEK293T cells. Pearson’s correlation coefficient (R) = 0.81.
