Comparative analysis of retroviral vector expression in mouse embryonal carcinoma cells.
A series of replication-defective retroviral vectors were assessed for their ability to efficiently transfer functional genes into undifferentiated cells. In these vectors (designated handicapped because of a deletion of enhancer and promoter sequences in the viral long terminal repeat) transcription of inserted genes is under the control of internal promoters. Although a composite promoter composed of a mutant polyoma virus enhancer (PyF441) coupled to the herpes simplex virus thymidine kinase promoter was anticipated to function efficiently, it was found to be significantly inferior to the mouse X-chromosome phosphoglycerate kinase (pgk-1) promoter, in its ability to express the selectable neomycin phosphotransferase gene in mouse embryonal carcinoma cells. The pHMB vector, which contains the pgk-1 promoter, was shown to confer the drug-resistant phenotype at high frequencies to F9 and P19 cells. This vector might prove to be of general utility for efficient gene expression in other developmental contexts.
Hawley
T S
TS
Department of Experimental Oncology, Ottawa Regional Cancer Centre, Ontario, Canada.
Sabourin
L A
LA
Hawley
R G
RG
D000818:Animals; D004352:Drug Resistance, Microbial / Q000235:genetics; D054278:Embryonal Carcinoma Stem Cells; D015870:Gene Expression; D005822:Genetic Vectors; D051379:Mice; D014411:Neoplastic Stem Cells; D010735:Phosphoglycerate Kinase / Q000235:genetics*; D010957:Plasmids; D011120:Polyomavirus / Q000235:genetics; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics*; D018139:Simplexvirus / Q000235:genetics; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D010735:Phosphoglycerate Kinase