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Transfer of genes into hematopoietic cells using recombinant DNA viruses
- Source :
- Proceedings of the National Academy of Sciences. 82:158-162
- Publication Year :
- 1985
- Publisher :
- Proceedings of the National Academy of Sciences, 1985.
-
Abstract
- The ability of recombinant DNA viruses to transfer genes into hematopoietic cells has been explored. A recombinant simian virus 40 (SV40) in which the early region had been replaced with the chloramphenicol acetyltransferase (CAT) gene driven by the promoter from Rous sarcoma virus (RSV), was constructed. This virus transferred the CAT gene more efficiently into mouse and human bone marrow cells and into the K562, MEL, and WEHI hematopoietic tissue culture cell lines, than the classical calcium phosphate DNA transfer procedure, as shown by assay for CAT activity 48 hr after infection. Recombinant SV40 virions were also shown to be capable of stably transforming Chinese hamster ovary cells by use of an early region recombinant containing the methotrexate-resistant dihydrofolate reductase (DHFR) gene driven by the RSV promoter. The entire DHFR transcriptional unit could be detected in the genome of transformed cells that were also shown to be resistant to methotrexate. A recombinant adenovirus stock containing the neomycin-resistance gene driven by the SV40 early promoter was used to infect the K562 and MEL hematopoietic cell lines to resistance to the antibiotic G418. Transformation frequency was 10- to 100-fold higher than that obtained with calcium phosphate-precipitated DNA. Most or all of the recombinant adenovirus genome was integrated as 1-3 copies in the transformed cells. These studies show the feasibility of using DNA viruses for introduction of new genetic material into hematopoietic cells.
- Subjects :
- Chloramphenicol O-Acetyltransferase
viruses
Genetic Vectors
Simian virus 40
medicine.disease_cause
Adenoviridae
Cell Line
law.invention
Mice
Acetyltransferases
law
Chlorocebus aethiops
Cell Adhesion
medicine
Animals
Gene
Rous sarcoma virus
Multidisciplinary
biology
Adenovirus genome
Chinese hamster ovary cell
Hematopoietic Tissue
Cell Transformation, Viral
Hematopoietic Stem Cells
biology.organism_classification
Virology
Molecular biology
Tetrahydrofolate Dehydrogenase
Gene Expression Regulation
Cell culture
DNA, Viral
Recombinant DNA
Genetic Engineering
Research Article
Subjects
Details
- ISSN :
- 10916490 and 00278424
- Volume :
- 82
- Database :
- OpenAIRE
- Journal :
- Proceedings of the National Academy of Sciences
- Accession number :
- edsair.doi.dedup.....7f6789b7c4d6ffc853cfbee88269dae8