A colony-forming assay for determining the establishment efficiency of S/MAR-containing nonviral episomal expression vectors.
A colony-forming assay for determining the establishment efficiency of S/MAR-containing nonviral episomal expression vectors.
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DOI：
10.1101/pdb.prot069500
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发表时间：
2012-06-01
影响因子：
--
通讯作者：
Lipps, Hans J
中科院分区：
文献类型：
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作者：
Hagedorn, Claudia;Baiker, Armin;Lipps, Hans J
与所有真核复制子一样，S/MAR（支架/基质附着区）载体的稳定建立是一个随机事件，取决于知之甚少的表观遗传因素，如染色质结构和核定位。建立效率描述了其中特定S/MAR载体在初始选择期后作为附加体稳定保留的细胞的百分比。S/MAR载体的预期建立效率为1- 5%。本文介绍了一种集落形成试验，可用于确定建立效率或产生单细胞克隆。
As with all eukaryotic replicons, the stable establishment of S/MAR (scaffold/matrix attached region) vectors is a stochastic event that depends on poorly understood epigenetic factors such as chromatin structure and nuclear localization. Establishment efficiency describes the percentage of cells in which a particular S/MAR vector is stably retained as an episome after an initial selection period. Expected establishment efficiency for S/MAR vectors is 1-5%. This article describes a colony-forming assay that may be used either to determine establishment efficiency or to generate single cell clones.