title: CRISPR/Cas9-generated p47phox-deficient cell line for Chronic Granulomatous Disease gene therapy vector development.
abstract: Development of gene therapy vectors requires cellular models reflecting the genetic background of a disease thus allowing for robust preclinical vector testing. For human p47phox-deficient chronic granulomatous disease (CGD) vector testing we generated a cellular model using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 to introduce a GT-dinucleotide deletion (ΔGT) mutation in p47phoxencoding NCF1 gene in the human acute myeloid leukemia PLB-985 cell line. CGD is a group of hereditary immunodeficiencies characterized by impaired respiratory burst activity in phagocytes due to a defective phagocytic nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. In Western countries autosomal-recessive p47phox-subunit deficiency represents the second largest CGD patient cohort with unique genetics, as the vast majority of p47phoxCGD patients carries ΔGT deletion in exon two of the NCF1 gene. The established PLB-985 NCF1 ΔGT cell line reflects the most frequent form of p47phox-deficient CGD genetically and functionally. It can be differentiated to granulocytes efficiently, what creates an attractive alternative to currently used iPSC models for rapid testing of novel gene therapy approaches.
authors.affiliation: Division of Immunology, University Children's Hospital Zurich, Zurich, Switzerland.|Children's Research Center, Zurich, Switzerland.|University of Zurich, Zurich, Switzerland.
authors.forename: Dominik
authors.initials: D
authors.lastname: Wrona
authors.affiliation: Division of Immunology, University Children's Hospital Zurich, Zurich, Switzerland.|Children's Research Center, Zurich, Switzerland.|University of Zurich, Zurich, Switzerland.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Siler
authors.affiliation: Division of Immunology, University Children's Hospital Zurich, Zurich, Switzerland.|Children's Research Center, Zurich, Switzerland.|University of Zurich, Zurich, Switzerland.
authors.forename: Janine
authors.initials: J
authors.lastname: Reichenbach
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006105:Granulomatous Disease, Chronic / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D018922:HL-60 Cells; D006801:Humans; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion
chemical_list: D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1