Recessive Dystrophic Epidermolysis Bullosa (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 C7, the major component of anchoring fibrils (AFs) which form key attachment structures for dermal-epidermal adherence. Here, we achieved highly efficient COL7A1 editing in primary RDEB keratinocytes (RDEB-K) and fibroblasts (RDEB-F) from two patients homozygous for the c.6508C>T (p.Gln2170*) mutation through CRISPR/Cas9-mediated Homology-Directed Repair. Three gRNAs targeting the c.6508C>T mutation or harboring sequences, were delivered together with hfCas9 as a ribonucleoprotein complex (RNP). Among them, one achieved 73% cleavage activity in primary RDEB-K and RDEB-F. Then, we treated RDEB-K and RDEB-F with this specific RNP and the corresponding donor template delivered as ssODN and achieved up to 58% of genetic correction as well as C7 rescue. Finally, grafting of corrected 3D skin onto nude mice induced re-expression and normal localization of C7 as well as AFs formation at the DEJ at five and ten weeks post-grafting. With this promising non-viral approach, we achieved therapeutically relevant specific gene editing which could be applicable to all mutations in exon 80 of COL7A1 in primary RDEB cells.