Gene-edited pseudogene resurrection corrects p47 phox -deficient chronic granulomatous disease

Pseudogenes are duplicated genes with mutations rendering them nonfunctional. For single-gene disorders with homologous pseudogenes, the pseudogene might be a target for genetic correction. Autosomal-recessive p47 -deficient chronic granulomatous disease (p47-CGD) is a life-threatening immune deficiency caused by mutations in , a gene with 2 pseudogenes, and . The most common mutation, a GT deletion (ΔGT) at the start of exon 2 (>90% of alleles), is constitutive to and . ΔGT results in premature termination, undetectable protein expression, and defective production of antimicrobial superoxide in neutrophils. We examined strategies for p47-CGD gene correction using engineered zinc-finger nucleases targeting the exon 2 ΔGT in induced pluripotent stem cells or CD34 hematopoietic stem cells derived from p47-CGD patients. Correction of ΔGT in pseudogenes restores oxidase function in p47-CGD, providing the first demonstration that targeted restoration of pseudogene function can correct a monogenic disorder.