TALEN-based gene correction for epidermolysis bullosa.

[dystrophic epidermolysis bullosa]

Recessive dystrophic epidermolysis bullosa (RDEB ) is characterized by a functional deficit of type VII collagen protein due to gene defects in the type VII collagen gene (COL 7 A 1 ). Gene augmentation therapies are promising , but run the risk of insertional mutagenesis . To abrogate this risk , we explored the possibility of using engineered transcription activator-like effector nucleases (TALEN ) for precise genome editing . We report the ability of TALEN to induce site-specific double -stranded DNA breaks (DSBs ) leading to homology-directed repair (HDR ) from an exogenous donor template . This process resulted in COL 7 A 1 gene mutation correction in primary fibroblasts that were subsequently reprogrammed into inducible pluripotent stem cells and showed normal protein expression and deposition in a teratoma -based skin model in vivo . Deep sequencing-based genome-wide screening established a safety profile showing on-target activity and three off-target (OT ) loci that , importantly , were at least 10 â€‰kb from a coding sequence . This study provides proof-of-concept for TALEN-mediated in situ correction of an endogenous patient-specific gene mutation and used an unbiased screen for comprehensive TALEN target mapping that will cooperatively facilitate translational application .

Diseases
Validation

Diseases presenting "primary fibroblasts" symptom

dystrophic epidermolysis bullosa

gm1 gangliosidosis

werner syndrome

x-linked adrenoleukodystrophy

You can validate or delete this automatically detected symptom