Retroviral mutants efficiently expressed in embryonal carcinoma cells.
The myeloproliferative sarcoma virus (MPSV) is a unique member of the Moloney murine sarcoma virus family. Due to mutations in the U3 region of its long terminal repeat, MPSV has an expanded host range that includes cells of the hematopoietic compartment. Using a MPSV recombinant containing the gene for neomycin-resistance (NeoR-MPSV), we demonstrate that the host range of MPSV also includes undifferentiated F9 embryonal carcinoma cells. Transfer of G418-resistance with NeoR-MPSV to F9 cells is almost as efficient as G418-resistance transfer to fibroblasts, in contrast to G418-resistance transfer to PCC4 embryonal carcinoma cells, which is at least 3 orders of magnitude lower. To isolate NeoR-MPSV mutants that are efficiently expressed in PCC4 cells, G418-resistant PCC4 cell lines were induced to differentiate, and the provirus was rescued by superinfection with murine leukemia virus. Viral isolates (PCMV-5 and -6; PCMV = PCC4 cell-passaged NeoR-MPSV) were obtained and assayed for expression in embryonal carcinoma cells. The efficiency of NeoR transfer was equally as high in both F9 and PCC4 as in fibroblasts. mos oncogene expression was unaltered as judged by transformation capability. No gross alteration in the coding region and in the long terminal repeat was detectable by restriction enzyme analysis. NeoR-MPSV and its mutants PCMV-5 and -6 can thus be utilized as vectors for the efficient transduction of genes into embryonic cells.
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004351:Drug Resistance; D005786:Gene Expression Regulation; D051379:Mice; D008980:Moloney murine sarcoma virus / Q000235:genetics*; D009154:Mutation; D009355:Neomycin / Q000494:pharmacology; D051381:Rats; D009053:Sarcoma Viruses, Murine / Q000235:genetics*; D013724:Teratoma / Q000235:genetics* / Q000473:pathology; D014161:Transduction, Genetic; D014212:Tretinoin / Q000494:pharmacology
D014212:Tretinoin; D009355:Neomycin