Figure 4.

(a) Sequencing diagram of amplified calpain-3 cDNA after genome editing, proves precise fusion between exon3 and exon4 and the restoration to WT CAPN3 sequence. (b) FACS gating strategy to distinguish/acquire cells with bi-allelic modifications (tdTomato and eGFP double-positive population). Lower left panel: immunofluorescence picture of an iPSC colony after the double-positive sorting. (c) FACS gating strategy to distinguish/acquire cells with t removal of the selection cassette (tdTomato and eGFP double-negative population). Lower left panel: PCR of verification of cassette excision at CAPN3 locus and chimeric Exon3/4 generation on the level of genomic DNA (lane “Isogenic LGMD2A”; PCR control lane “patient LGMD2A”).