879 Development of a method identifying revertant mosaicism in recessive dystrophic epidermolysis bullosa using long-read sequencing

Recessive dystrophic epidermolysis bullosa (RDEB) is a cutaneous genetic disease, which characterized by the loss of functional type VII collagen. The epidermis becomes extremely fragile without collagen, and patients suffer from repetitive blistering scar formation. RDEB caused mutations in COL7A1 gene (coding collagen), most have two different on each gene. Interestingly, it known that part skin can be reverted to normal appearance; this phenomenon called revertant mosaicism. One possible mechanism inducing mosaicism chromosome crossover are gathered one chromosome, resulting recovery other chromosome. Revertant mosaicism, also referred as natural therapy, has attracted much attention for elucidating pathogenesis developing treatment disease. However, been technically challenging identify whether pathogenic same difficulty barrier field. In study, we report an efficient method using long-read sequencing analyze We improved existing method, nCATS, uses CRISPR/Cas9 technology enrich target region genome, barcoded adapters introducing pre-amplification step. believe will useful not only but analysis many diseases.