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. 2023 Oct 24;14(2):362–379. doi: 10.1158/2159-8290.CD-23-0402

Figure 5.

Figure 5. Impact of correction of the mutant TP53 sequence to wt TP53 in the MDA-MB-231 and AU565 human cancer cell lines. A, Schematic of the components used for electroporation to correct the mutant TP53 gene sequence to wt TP53 in human cancer cells. The crRNA recognition site (underlined), PAM sequence (in green), and the cut site (dashed vertical line) for the MDA-MB-231 and AU565 mutTP53 target sequence are presented (K280 and H175 residues in yellow). Bottom, the ssDNA HDR donor sequences are presented: the modified codon (in red) together with the PAM sequence silent mutation (in orange) are shown. B, Percentages of genomes carrying the intended TP53 edit at the indicated time points. N = 3 independent experiments. Individual values of biological replicates their mean and SEM are reported. C, The mRNA abundance for the indicated genes at the indicated time points in the absence (-) or presence (+) of treatment with 5 μmol/L nutlin-3a for 12 hours. N = 3 independent experiments. The data are presented as the mean and SEM of three independent experiments.

Impact of correction of the mutant TP53 sequence to wt TP53 in the MDA-MB-231 and AU565 human cancer cell lines. A, Schematic of the components used for electroporation to correct the mutant TP53 gene sequence to wt TP53 in human cancer cells. The crRNA recognition site (underlined), PAM sequence (in green), and the cut site (dashed vertical line) for the MDA-MB-231 and AU565 mutTP53 target sequence are presented (K280 and H175 residues in yellow). Bottom, the ssDNA HDR donor sequences are presented: the modified codon (in red) together with the PAM sequence silent mutation (in orange) are shown. B, Percentages of genomes carrying the intended TP53 edit at the indicated time points. N = 3 independent experiments. Individual values of biological replicates their mean and SEM are reported. C, The mRNA abundance for the indicated genes at the indicated time points in the absence (-) or presence (+) of treatment with 5 μmol/L nutlin-3a for 12 hours. N = 3 independent experiments. The data are presented as the mean and SEM of three independent experiments.