EMBRYONIC STEM-CELL VIRUS, A RECOMBINANT MURINE RETROVIRUS WITH EXPRESSION IN EMBRYONIC STEM-CELLS
EMBRYONIC STEM-CELL VIRUS, A RECOMBINANT MURINE RETROVIRUS WITH EXPRESSION IN EMBRYONIC STEM-CELLS
复制标题
DOI：
10.1073/pnas.87.23.9202
复制
发表时间：
1990-12-01
影响因子：
11.1
通讯作者：
OSTERTAG, W
中科院分区：
文献类型：
--
作者：
GREZ, M;AKGUN, E;OSTERTAG, W
莫洛尼鼠白血病病毒及其衍生载体的表达仅限于未分化的小鼠胚胎癌和胚胎干（ES）细胞。我们开发了一种逆转录病毒载体，即鼠胚胎干细胞病毒 (MESV)，它在胚胎癌和 ES 细胞中具有活性。 MESV 源自一种逆转录病毒突变体 [PCC4 细胞传代的骨髓增殖肉瘤病毒 (PCMV)]，在胚胎癌细胞中表达，但在 ES 细胞中不表达。 PCMV 的增强子区域在两种细胞类型中均显示出功能，但发现 PCMV 5''非翻译区域内的序列限制 ES 细胞中的病毒表达。用从 dl-587rev 逆转录病毒获得的相关序列替换该区域会产生 MESV，这是一种改良的 PCMV 病毒，赋予 G418 对成纤维细胞和 ES 细胞的抗性，并具有相似的效率。 ES 细胞中 MESV 的表达是由病毒基因组 5'' 长末端重复序列内的转录调控元件介导的。
The expression of Moloney murine leukemia virus and vectors derived from it is restricted in undifferentiated mouse embryonal carcinoma and embryonal stem (ES) cells. We have developed a retroviral vector, the murine embryonic stem cell virus (MESV), that is active in embryonal carcinoma and ES cells. MESV was derived from a retroviral mutant [PCC4-cell-passaged myeloproliferative sarcoma virus (PCMV)] expressed in embryonal carcinoma cells but not in ES cells. The enhancer region of PCMV was shown to be functional in both cell types, but sequences within the 5'' untranslated region of PCMV were found to restrict viral expression in ES cells. Replacement of this region by related sequences obtained from the dl-587rev retrovirus results in MESV, a modified PCMV virus that confers G418 resistance to fibroblasts and ES cells with similar efficiencies. Expression of MESV in ES cells is mediated by transcriptional regulatory elements within the 5'' long terminal repeat of the viral genome.