Cloning and expression of the cDNA encoding rat granulocyte colony-stimulating factor
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Elsevier
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Granulocyte colony-stimulating factor (G-CSF) acts on precursor hematopoietic cells to control the production and maintenance of neutrophils. Recombinant G-CSF (re-G-CSF)is used clinically to treat patients with neutropenia and has greatly reduced the infection risk associated with bone marrow transplantation. Cyclic hematopoiesis, a stem cell defect characterized by severe recurrent neutropenia, occurs in man and grey collie dogs, and can be treated by administration of re-G-CSF. Availability of the rat G-CSF cDNA would benefit the use of rats as models of gene therapy for the treatment of cyclic hematopoiesis. In preliminary rat experiments, retroviral-mediated expression of canine G-CSF caused neutralizing antibody formation which precluded long-term increases in neutrophil counts. To overcome this problem we cloned the rat G-CSF cDNA from RNA isolated from skin fibroblasts. The rat G-CSF sequence shared a high degree of identity in both the coding and non-coding regions with both the murine G-CSF (85%) and human G-CSF (74%). The signal peptides of murine and human G-CSF both contained 30 amino acids (aa), whereas the deduced signal sequence for rat G-CSF possessed 21 aa. A retrovirus encoding the rat G-CSF cDNA synthesized bioactive G-CSF from transduced vascular smooth muscle cells.
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Gene therapy , Neutropenia , Nucleotide sequence , complementary dna , granulocyte colony stimulating factor , neutralizing antibody , recombinant granulocyte colony stimulating factor , animal cell , animal experiment , animal model , bone marrow transplantation , dog , gene therapy , graft infection , hematopoiesis , hematopoietic cell , human , human cell , molecular cloning , neutropenia , neutrophil , nonhuman , priority journal , rat , retrovirus , stem cell , vascular smooth muscle , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , DNA, Complementary , Genetic Vectors , Granulocyte Colony-Stimulating Factor , Mice , Molecular Sequence Data , Rats , Sequence Analysis, DNA , Sequence Homology , Animalia , Canis familiaris , Murinae , unidentified retrovirus
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Citação
Gene, v. 175, n. 1-2, p. 101-104, 1996.
