MAR characteristic motifs mediate episomal vector in CHO cells
MAR characteristic motifs mediate episomal vector in CHO cells
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MAR 特征基序介导 CHO 细胞中的附加型载体
DOI：
10.1016/j.gene.2015.01.032
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发表时间：
2015-04-01
期刊：
影响因子：
3.5
通讯作者：
Yang, Xianjun
中科院分区：
文献类型：
--
作者：
Lin, Yan;Li, Zhaoxi;Yang, Xianjun
理想的基因治疗载体应该能够实现持续的转基因表达，而不受安全性和可重复性的限制。最近的研究对染色体基质附着区（MAR）介导遗传元件的附加型维持的能力的洞察允许环状附加型载体的发展。虽然MAR介导的工程载体已经开发出来，但MAR的哪些基序在与宿主基因组相互作用期间赋予这种功能还知之甚少。在这里，我们报告了一个人工合成的DNA片段，只含有特征性的基序序列，作为替代人β-干扰素基质附着区序列。研究了该载体在CHO细胞中介导基因转移的潜力。发现短的合成MAR基序以低拷贝数介导附加型载体许多代而不整合到宿主基因组中。较高的转基因表达维持至少4个月。此外，MAR保持游离型，并赋予持续的EGFP表达，即使在非选择性CHO细胞。所有结果表明，MAR特征序列载体可以作为稳定的附加体在CHO细胞中发挥作用，支持长期有效的转基因表达。(C)2015 Elsevier B. V.版权所有。
An ideal gene therapy vector should enable persistent transgene expression without limitations in safety and reproducibility. Recent researches' insight into the ability of chromosomal matrix attachment regions (MARs) to mediate episomal maintenance of genetic elements allowed the development of a circular episomal vector. Although a MAR-mediated engineered vector has been developed, little is known on which motifs of MAR confer this function during interaction with the host genome. Here, we report an artificially synthesized DNA fragment containing only characteristic motif sequences that served as an alternative to human beta-interferon matrix attachment region sequence. The potential of the vector to mediate gene transfer in CHO cells was investigated. The short synthetic MAR motifs were found to mediate episomal vector at a low copy number for many generations without integration into the host genome. Higher transgene expression was maintained for at least 4 months. In addition, MAR was maintained episomally and conferred sustained EGFP expression even in nonselective CHO cells. All the results demonstrated that MAR characteristic sequence-based vector can function as stable episomes in CHO cells, supporting long-term and effective transgene expression. (C) 2015 Elsevier B.V. All rights reserved.