Rous sarcoma virus is integrated but not expressed in chicken early embryonic cells.
We have developed a protocol that allows us to infect chicken early embryonic (CEE) cells with high efficiency. This was achieved by exposing the CEE cells to a semicontinuous dose of Rous sarcoma virus (RSV) for a period of 20 hr. Southern blot analysis indicated that an average of one proviral copy is integrated per embryonic cell. However, there was no production of infectious viral particles by the cells containing the proviral genome, although low levels of full-length genomic RNA could be detected by RNA transfer blot analysis. These low RNA levels contrast with the 100- to 1000-fold higher levels found in RSV-infected chicken embryo fibroblasts. We conclude that in cells derived from pregastrulating chicken embryos, RSV DNA is integrated into the cell genome but fails to be expressed in an efficient manner. These primary cells can therefore be used to identify factors involved in regulation of retroviral gene expression in normal cells. Such factors may also be instrumental in elucidating basic mechanisms involved in gene regulation during early development in higher vertebrates.
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics*; D001756:Blastoderm / Q000166:cytology; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D002642:Chick Embryo; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification
D004279:DNA, Viral; D012367:RNA, Viral