Gene targeting of X chromosome-linked chronic granulomatous disease locus in a human myeloid leukemia cell line and rescue by expression of recombinant gp91[sup phox]

Journal Article · Mon Nov 15 04:00:00 EST 1993 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.90.21.9832· OSTI ID:7105204

Zhen, Ling; Dinauer, M C ^[1]; King, A A.J.; Xiao, Yonghong ^[2]; Chanock, S J ^[3]; Orkin, S H ^[4]

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1. Indiana Univ. Medical Center, Indianapolis, IN (United States)
2. Children's Hospital and the Dana Farber Cancer Institute, Harvard Medical School, Boston, MA (United States)
3. Children's Hospital and the Dana Farber Cancer Institute, Harvard Medical School, Boston, MA (United States) National Cancer Institute, Bethesda, MD (United States)
4. Children's Hospital and the Dana Farber Cancer Institute, Harvard Medical School, Boston, MA (United States) Howard Hughes Medical Institute, Boston, MA (United States)

The X chromosome-linked chronic granulomatous disease (X-CGD) locus, which encodes the gp91[sup phox] subunit of the phagocyte respiratory-burst oxidase cytochrome b, was disrupted by homologous recombination in the PLB-985 human myeloid cell line to develop an in vitro model of XCGD. Superoxide formation was absent in targeted cells after differentiation of granulocytes but was rescued by stable transfection and expression of wild-type gp91[sup phox] cDNA. The targeted cell line should be useful in experiments aimed at defining functional regions within gp91[sup phox] by expression of mutant gp91[sup phox] cDNAs, complementing studies of naturally occurring mutations in X-CGD. In addition, the mutant line provides a model system in which to establish an experimental basis for the treatment of X-CGD patients with gene replacement therapy. Rescued clones containing even modest amounts of recombinant gp91[sup phox] had respiratory-burst activity comparable to the wild-type PLB-985 line, suggesting that functional correction of X-CGD neutrophils may not require high-level expression of gp91[sup phox].

• OSTI ID: 7105204
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 90:22; ISSN PNASA6; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
BIOLOGICAL MODELS
CHROMOSOMES
DISEASES
ENZYMES
GENE MUTATIONS
GENETIC MAPPING
HEREDITARY DISEASES
HETEROCHROMOSOMES
HUMAN CHROMOSOMES
HUMAN X CHROMOSOME
MAPPING
MUTATIONS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PHAGOCYTES
PROTEINS
SOMATIC CELLS
X CHROMOSOME