Expression of the human. beta. -globin gene following retroviral-mediated transfer into multipotential hematopoietic progenitors of mice

Journal Article · Mon Aug 01 00:00:00 EDT 1988 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.85.16.6062· OSTI ID:5513152

Karlsson, S; Bodine, D M; Perry, L; Papayannopoulou, T; Nienhuis, A W ^[1]

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1. National Institutes of Health, Bethesda, MD (USA)

Efficient transfer of the {beta}-globin gene into primitive hematopoietic progenitors was achieved with consistent and significant expression in the progeny of those cells. Retroviral vectors containing the intact genomic human {beta}-globin gene and the neomycin (G418)-resistance (neo{sup R}) gene were constructed. These gave titers of 10{sup 6} or more neo{sup R} colony-forming units/ml when packaged in {psi}2 cells. Mouse bone marrow cells were infected by coculture with producer cells and injected into lethally irradiated animals. Several parameters were varied to enhance infection frequency of colony-forming units, spleen (CFU-S); overall 41% of 116 foci studied contained an intact proviral genome. The human {beta}-globin gene was expressed in 31 of 35 CFU-S-derived spleen colonies that contained the intact vector genome at levels ranging from 1% to 5% of that of the mouse {beta}-globin genes. Infected bone marrow cells were also injected into genetically anemic W/W{sup v} recipients without prior irradiation. Human {beta}-globin chains were detected in circulating erythrocytes by immunofluorescent staining with a specific monoclonal antibody. All animals injected with donor cells that had been cultured in G418 (1 mg/ml) for 48 hr after retroviral infection had circulating erythrocytes containing human {beta}-globin chains between 3 and 8 weeks after transplantation.

• OSTI ID: 5513152
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 85:16; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550401 -- Genetics-- Tracer Techniques
560152* -- Radiation Effects on Animals-- Animals
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
BONE MARROW CELLS
CFU
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
ELECTROPHORESIS
ERYTHROCYTES
EXTERNAL IRRADIATION
GENE RECOMBINATION
GENE REGULATION
GENES
GLOBIN
IRRADIATION
ISOTOPES
LETHAL IRRADIATION
LIGHT NUCLEI
MAMMALS
MAN
MATERIALS
MICE
MICROORGANISMS
MONOCLONAL ANTIBODIES
NUCLEI
ODD-ODD NUCLEI
PARASITES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PRIMATES
RADIOISOTOPES
RODENTS
SOMATIC CELLS
SPLEEN CELLS
TRANSCRIPTION
TRANSPLANTS
VERTEBRATES
VIRUSES
WHOLE-BODY IRRADIATION