Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells

Shortened nuclear matrix attachment regions are sufficient for replication and maintenance of episomes in mammalian cells
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缩短的核基质附着区域足以在哺乳动物细胞中复制和维持附加体

DOI:
10.1091/mbc.e19-02-0108
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发表时间:
2019-10
影响因子:
3.3
通讯作者:
Yi Dan Dan
Yi Dan Dan
中科院分区:
生物学3区
文献类型:
--
作者:
Wang Xiao Yin;Zhang Xi;Wang Tian Yun;Jia Yan Long;Xu Dan Hua;Yi Dan Dan

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相似文献

基质附着区(MARs)可以介导载体附加体在哺乳动物细胞中的复制;然而,其分子作用方式仍不清楚。在这里,我们评估了MAR的特点和机制,介导的附加型载体复制在哺乳动物细胞。克隆β-干扰素MAR片段的五个缩短的亚片段并转移到CHO细胞中,并测定转基因表达水平、基因的存在和附加体维持机制。三种缩短的MAR衍生物(位置781-1320、1201-1740和1621-2201)保留了完整的MAR活性并介导附加型载体复制。此外,与原质粒pEPI-1相比,三个截短的MAR具有更高的转基因表达水平、更高的集落形成效率和更持久的转基因表达,并且三个功能截短的MAR可以与SAF-A MAR结合蛋白结合。这些结果表明,缩短的MAR足以在CHO细胞中复制和维持附加体。
Matrix attachment regions (MARs) can mediate the replication of vector episomes in mammalian cells; however, the molecular mode of action remains unclear. Here, we assessed the characteristics of MARs and the mechanism that mediates episomal vector replication in mammalian cells. Five shortened subfragments of β-interferon MAR fragments were cloned and transferred into CHO cells, and transgene expression levels, presence of the gene, and the episomal maintenance mechanism were determined. Three shortened MAR derivatives (position 781–1320, 1201–1740, and 1621–2201) retained full MAR activity and mediated episomal vector replication. Moreover, the three shortened MARs showed higher transgene expression levels, greater efficiency in colony formation, and more persistent transgene expression compared with those of the original pEPI-1 plasmid, and three functional truncated MARs can bind to SAF-A MAR-binding protein. These results suggest that shortened MARs are sufficient for replication and maintenance of episomes in CHO cells.
DOI: 10.1038/mtna.2013.47
发表时间: 2013-09-03
期刊: Molecular therapy. Nucleic acids
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