MULTIPLE MODIFICATIONS IN CIS-ELEMENTS OF THE LONG TERMINAL REPEAT OF RETROVIRAL VECTORS LEAD TO INCREASED EXPRESSION AND DECREASED DNA METHYLATION IN EMBRYONIC CARCINOMA-CELLS
MULTIPLE MODIFICATIONS IN CIS-ELEMENTS OF THE LONG TERMINAL REPEAT OF RETROVIRAL VECTORS LEAD TO INCREASED EXPRESSION AND DECREASED DNA METHYLATION IN EMBRYONIC CARCINOMA-CELLS
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DOI：
10.1128/jvi.69.2.748-755.1995
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发表时间：
1995-02-01
影响因子：
5.4
通讯作者：
KOHN, DB
中科院分区：
文献类型：
--
作者：
CHALLITA, PM;SKELTON, D;KOHN, DB
小鼠逆转录病毒在胚胎癌（EC）和胚胎干细胞中的感染受到严格限制。莫洛尼鼠白血病病毒 (MoMuLV) 长末端重复序列 (LTR) 的转录单位在 EC 和胚胎干细胞中失活，导致原病毒甲基化增加。在本研究中，F9 EC 细胞中的表达是通过在基于 MoMuLV 的逆转录病毒载体中包含三种修饰的新型逆转录病毒载体实现的：存在骨髓增殖性肉瘤病毒 LTR、引物结合位点的替换以及 LTR 5' 端阴性对照区的删除或去甲基化序列的插入。我们得出的结论是，EC 细胞中 MoMuLV LTR 表达的抑制是通过影响原病毒甲基化状态的多个顺式作用元件的累加效应介导的。
Infection by murine retroviruses in embryonic carcinoma (EC) and embryonic stem cells is highly restricted. The transcriptional unit of the Moloney murine leukemic virus (MoMuLV) long terminal repeat (LTR) is inactive in EC and embryonic stem cells in association,vith increased proviral methylation. In this study, expression in F9 EC cells was achieved from novel retroviral vectors containing three modifications in the MoMuLV-based retroviral vector: presence of the myeloproliferative sarcoma virus LTR, substitution of the primer binding site, and either deletion of a negative control region at the 5' end of the LTR or insertion of a demethylating sequence. We conclude that inhibition of expression from the MoMuLV LTR in EC cells is mediated through the additive effects of multiple cis-acting elements affecting the state of methylation of the provirus.