Fig. 1
Gene editing-mediated introduction of the c.1054C > T mutation in the CLN3 locus alters splicing. a Representation of biallelic donors containing distinctive fluorescent positive selection modules (PSM) and the targeted genomic region. b FACS plots depicting the different populations through the steps of the editing process. The upper panel represents the line after the integration of the PSM and the lower panel, the enrichment of the line after several rounds of sorting for the double-positive population. Plots are accompanied in the right side by microscope images for proper visualization of each step. Scale bars, 200 μm. c FACS plots depicting the different populations through the steps of the editing process. The upper panel represents the line after the excision of the PSM and the lower panel, the purification of the line after several rounds of sorting for the double-negative population. Plots are accompanied in the right side by microscope images for proper visualization of each step. Scale bars, 200 μm. d Sanger sequencing chromatogram of the obtained polyclone highlighting the introduction of the mutation and the silent PAM modifications. e Representative RT-PCR gel showing the different band amplification sizes in the CLN3Q352X mutant hiPSCs and patient PBMCs, as compared to the controls. f Second band corresponds to an exon-skipping event of the mutated exon. Sanger sequencing confirmed the junction between neighboring exons