156 Targeted ex-vivo correction of COL7A1 through RNP-based CRISPR/Cas9 and Homology- Directed Repair to treat recessive dystrophic epidermolysis bullosa

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RDEB is a rare and severe genetic skin disease responsible for blistering of the skin and mucosa after minor trauma. RDEB is caused by a wide variety of mutations in COL7A1 encoding type VII collagen, the major component of anchoring fibrils critical attachment structures that adhere the epidermis to the dermis. Here, we achieved highly efficient COL7A1 editing in primary RDEB keratinocytes, fibroblasts, and 3D-skin grafts through CRISPR/Cas9-mediated Homology Directed Repair. First, we designed three guide RNAs (gRNAs) targeting the specific mutation or sequences in close distance to the c.6508C>T (p.Gln2170*) mutation to be corrected in exon 80.