Expression of the human. beta. -globin gene following retroviral-mediated transfer into multipotential hematopoietic progenitors of mice
- 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), Vol. 85:16; ISSN 0027-8424
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
BONE MARROW CELLS
GENE RECOMBINATION
GLOBIN
GENE REGULATION
MICE
LETHAL IRRADIATION
CFU
ELECTROPHORESIS
ERYTHROCYTES
GENES
MAN
MONOCLONAL ANTIBODIES
PHOSPHORUS 32
SPLEEN CELLS
TRANSCRIPTION
TRANSPLANTS
VIRUSES
WHOLE-BODY IRRADIATION
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
EXTERNAL IRRADIATION
IRRADIATION
ISOTOPES
LIGHT NUCLEI
MAMMALS
MATERIALS
MICROORGANISMS
NUCLEI
ODD-ODD NUCLEI
PARASITES
PHOSPHORUS ISOTOPES
PRIMATES
RADIOISOTOPES
RODENTS
SOMATIC CELLS
VERTEBRATES
560152* - Radiation Effects on Animals- Animals
550401 - Genetics- Tracer Techniques