Retrovirus-mediated gene transfer into embryonal carcinoma and hemopoietic stem cells: Expression from a hybrid long terminal repeat
article
Retroviral vectors can be used as an efficient gene delivery system in a wide variety of cell types. However, in some cell types, such as embyronal carcinoma (EC) cells or normal bone-marrow cells the expression of genes introduced by retroviral vectors has been very inefficient. This expression block has severely hampered the application of retroviral vector systems in those cell types. The enhancer sequences present in the long terminal repeat (LTR) of retroviruses are known to be responsible for the tissue specificity of viral expression. Therefore, we set out to construct a vector in which this enhancer element has been replaced. A recombinant retrovirus was constructed in which the enhancer from the Moloney murine leukemia virus LTR was replaced by the enhancer of a mutant polyoma virus (PyF101) that was selected to grow on EC cells. A neomycin-resistance marker (neo(R)) was placed under the transcriptional control of the hybrid LTR. Following infection with this virus, neo(R) was expressed in EC cells, as well as in the hemopoietic progenitor cells present in normal murine bone marrow. Moreover, upon transplantation of infected bone marrow cells into lethally irradiated mice, neo(R) expression was sustained in hemopoietic cells of the engrafted recipients.
Chemicals/CAS: Genetic Vectors; Neomycin, 1404-04-2
Chemicals/CAS: Genetic Vectors; Neomycin, 1404-04-2
Topics
Enhancergene therapyhemopoietic stem cellsMoloney murine leukemia viruspolyoma F101recombinant DNAretroviral vectorsanimal cellarticleembryonal carcinomagene therapygene transfergenetic engineeringhereditymousenonhumanpriority journalretrovirusstem cellAnimalBone Marrow DiseasesBone Marrow TransplantationCells, CulturedDrug Resistance, MicrobialEnhancer Elements (Genetics)Gene ExpressionGenes, ViralGenetic VectorsHematopoietic Stem CellsMiceMice, Inbred C57BLMice, Inbred CBANeomycinProvirusesRepetitive Sequences, Nucleic AcidRetroviridaeTransfectionTumor Stem CellsVirus CultivationAnimaliaMoloney murine leukemia virusMurinaePolyomavirusunidentified retrovirus
TNO Identifier
230969
ISSN
03781119
Source
Gene, 84(2), pp. 419-427.
Pages
419-427
Files
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