Gene-edited pseudogene resurrection corrects p47phox-deficient chronic granulomatous disease

• Published in: Blood advances 2017-01, Vol.1 (4), p.270-278
• Main Authors: Merling, Randall K., Kuhns, Douglas B., Sweeney, Colin L., Wu, Xiaolin, Burkett, Sandra, Chu, Jessica, Lee, Janet, Koontz, Sherry, Di Pasquale, Giovanni, Afione, Sandra A., Chiorini, John A., Kang, Elizabeth M., Choi, Uimook, De Ravin, Suk See, Malech, Harry L.
• Format: Article
• Language: English
• Subjects: Hematopoiesis and Stem Cells
• ISSN: 2473-9529; 2473-9537
• DOI: 10.1182/bloodadvances.2016001214

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 p47phox-deficient chronic granulomatous disease (p47-CGD) is a life-threatening immune deficiency caused by mutations in NCF1, a gene with 2 pseudogenes, NCF1B and NCF1C. The most common NCF1 mutation, a GT deletion (ΔGT) at the start of exon 2 (>90% of alleles), is constitutive to NCF1B and NCF1C. NCF1 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 NCF1 pseudogenes restores oxidase function in p47-CGD, providing the first demonstration that targeted restoration of pseudogene function can correct a monogenic disorder. •Gene-editing correction of the GT deletion in exon 2 of NCF1 pseudogenes corrects p47phox-deficient chronic granulomatous disease.•The nonfunctional pseudogenes NCF1B and NCF1C can be resurrected to produce functional p47phox protein by gene editing.