CRISPR/Cas9-based genetic correction for recessive dystrophic epidermolysis bullosa

Recessive dystrophic epidermolysis bullosa (RDEB) is a severe disorder caused by mutations to the COL7A1 gene that deactivate production of a structural protein essential for skin integrity. Haematopoietic cell transplantation can ameliorate some of the symptoms; however, significant side effects from the allogeneic transplant procedure can occur and unresponsive areas of blistering persist. Therefore, we employed genome editing in patient-derived cells to create an autologous platform for multilineage engineering of therapeutic cell types. The clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 system facilitated correction of an RDEB-causing COL7A1 mutation in primary fibroblasts that were then used to derive induced pluripotent stem cells (iPSCs). The resulting iPSCs were subsequently re-differentiated into keratinocytes, mesenchymal stem cells (MSCs) and haematopoietic progenitor cells using defined differentiation strategies. Gene-corrected keratinocytes exhibited characteristic epithelial morphology and expressed keratinocyte-specific genes and transcription factors. iPSC-derived MSCs exhibited a spindle morphology and expression of CD73, CD90 and CD105 with the ability to undergo adipogenic, chondrogenic and osteogenic differentiation in vitro in a manner indistinguishable from bone marrow-derived MSCs. Finally, we used a vascular induction strategy to generate potent definitive haematopoietic progenitors capable of multilineage differentiation in methylcellulose-based assays. In totality, we have shown that CRISPR/Cas9 is an adaptable gene-editing strategy that can be coupled with iPSC technology to produce multiple gene-corrected autologous cell types with therapeutic potential for RDEB. Epidermolysis bullosa: Gene-editing offers fix for skin condition Gene-editing coupled with patient-specific stem cell technology could help treat the rare skin disorder epidermolysis bullosa (EB). A team led by Jakub Tolar from the University of Minnesota, Minneapolis, USA, took skin punch biopsies from children with a recessive form of EB caused by mutations in the type VII collagen gene COL7A1 . These children have extremely fragile skin that blisters and tears from even the smallest trauma. The researchers used CRISPR/Cas9 gene-editing to correct the mutations in fibroblast cells from the skin. They then reprogrammed the gene-corrected cells into induced pluripotent stem cells, before coaxing the stems cells to form functional outer skin cells, b