302 Availability of mRNA Obtained from Peripheral Blood Mononuclear Cells for Mutational Analysis in Dystrophic Epidermolysis Bullosa

Dystrophic epidermolysis bullosa (DEB) is an inheritable blistering disease caused by mutations in COL7A1, which encodes type VII collagen. To address the issue of genotype-phenotype correlations DEB, analyzing consequences COL7A1 using mRNA indispensable. Herein we established a novel method for testing effect DEB extracted from peripheral blood mononuclear cells (PBMCs). We investigated four (c.6573 + 1G > C, c.6216 5G T, c.7270C T and c.2527C T) three Japanese individuals with recessive DEB. The detected two recurrent mutation (c.7270C accurately. In addition, it aberrant splicing resulting (c.2527C was previously reported as nonsense mutation. Furthermore, revealed that collagen-expressing PBMCs have similar cell surface markers mesenchymal stem cells; they were CD105+, CD29+, CD45-, CD34-, suggesting small number or stromal are circulating blood, enables us to detect PBMCs. Taken together, our obtained will significantly contribute genetic diagnoses therapies