Design of retrovirus vectors for transfer and expression of the human. beta. -globin gene
Regulated expression of the human ..beta..-globin gene has been demonstrated in cultured murine erythroleukemia cells and in mice after retrovirus-mediated gene transfer. However, the low titer of recombinant viruses described to date results in relatively inefficient gene transfer, which limits their usefulness for animal studies and for potential gene therapy in humans for diseases involving defective ..beta..-globin genes. The authors found regions that interfered with virus production within intron 2 of the ..beta..-globin gene and on both sides of the gene. The flanking regions could be removed, but intron 2 was required for ..beta..-globin expression. Inclusion of ..beta..-globin introns necessitates an antisense orientation of the gene within the retrovirus vector. However, they found no effect of the antisense ..beta..-globin transcription on virus production. A region downstream of the ..beta..-globin gene that stimulates expression of the gene in transgenic mice was included in the viruses without detrimental effects on virus titer. Virus titers of over 10/sup 6/ CFU/ml were obtained with the final vector design, which retained the ability to direct regulated expression of human ..beta..-globin in murine erythroleukemia cells. The vector also allowed transfer and expression of the human ..beta..-globin gene in hematopoietic cells (CFU-S cells) in mice.
- Research Organization:
- Fred Hutchinson Cancer Research Center, Seattle, WA (USA)
- OSTI ID:
- 6246645
- Journal Information:
- J. Virol.; (United States), Vol. 62:11
- Country of Publication:
- United States
- Language:
- English
Similar Records
A majority of mice show long-term expression of a human. beta. -globin gene after retrovirus transfer into hematopoietic stem cells
Retrovirus transduction: Segregation of the viral transforming function and the Herpes Simplex virus tk gene in infectious friend spleen focus-forming virus thymidine kinase vectors
Related Subjects
GLOBIN
GENE REGULATION
VIRUSES
GENE RECOMBINATION
BONE MARROW CELLS
CELL CULTURES
GENES
MAN
MICE
TRANSCRIPTION
TUMOR CELLS
ANIMAL CELLS
ANIMALS
CONNECTIVE TISSUE CELLS
MAMMALS
MICROORGANISMS
ORGANIC COMPOUNDS
PARASITES
PRIMATES
PROTEINS
RODENTS
SOMATIC CELLS
VERTEBRATES
550200* - Biochemistry