Han, Sang Won [UNESP]Ramesh, N.Osborne, William R.A.2014-05-272014-05-271996-10-10Gene, v. 175, n. 1-2, p. 101-104, 1996.0378-1119http://hdl.handle.net/11449/64860Granulocyte 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.101-104engGene therapyNeutropeniaNucleotide sequencecomplementary dnagranulocyte colony stimulating factorneutralizing antibodyrecombinant granulocyte colony stimulating factoranimal cellanimal experimentanimal modelbone marrow transplantationdoggene therapygraft infectionhematopoiesishematopoietic cellhumanhuman cellmolecular cloningneutropenianeutrophilnonhumanpriority journalratretrovirusstem cellvascular smooth muscleAmino Acid SequenceAnimalsBase SequenceCloning, MolecularDNA, ComplementaryGenetic VectorsGranulocyte Colony-Stimulating FactorMiceMolecular Sequence DataRatsSequence Analysis, DNASequence HomologyAnimaliaCanis familiarisMurinaeunidentified retrovirusArtigo10.1016/0378-1119(96)00131-XAcesso restrito2-s2.0-0030579145