Fig. 5: Evaluation of eCas12f1 as a potential cancer therapy tool.

a Schematic illustration of sgRNA designed to disrupt exon3 of the PLK1 gene. Underline, sgRNA spacer sequence; light blue letters, PAM sequence. b Indel frequency of eCas12f1-PLK1 in SKBR-3 cells. 36.69% of indel frequencies were observed in GFP-positive SKBR-3 cells. NTC, non-targeting control. c Images of SKBR-3 colonies stained with crystal violet for the colony formation assay. d Quantification of the relative colony formation ability. e Schematic illustration of sgRNA designed to disrupt exon15 of the BRAF with V600E mutation. Underline, sgRNA spacer sequence; light blue letters, PAM sequence; light green letters, wild type base pair in BRAF; red letters, base pair mutation in BRAF; black box, amino acid code. f, g Indel frequency of eCas12f1-BRAF in HEK293T cells (f) and A375 cells (g). 18.97% of indel frequencies were observed only in GFP-positive A375 cells. NTC, non-targeting control. h, i Images of HEK293T (h) and A375 (i) colonies stained with crystal violet for the colony formation assay after eCas12f1 treatment. j, k Quantification of the relative colony formation ability for HEK293T (j) and A375 (k) cells. 69.67% reduction in cell proliferation was observed only in A375 cells treated with eCas12f1 targeting BRAF V600E mutation gene. Data in the bar graphs represent mean ± s.d. of three independent biological replicates. P-values were obtained using the two-tailed Student’s t-test. ns, not significant. Source data are provided as a Source Data file.