Here, we outlined an allele-specific CRISPR-Cas9-based gene-editing approach that is able to disrupt specifically the KRT14 mutant gene and fully restore functional intermediate filaments in epidermal stem cells cultivated from an EBS patient carrying a de novo monoallelic c.475/495del21 dominant mutation in exon 1 of KRT14. This evidence concerns the gene KRT14 and epidermolysis bullosa simplex.