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人工弯曲 DNA 作为转录激活物在小鼠胚胎干细胞中的活性。

Competence of an artificial bent DNA as a transcriptional activator in mouse ES cells.

机构信息

Department of Biology, Faculty of Education and Integrated Arts and Sciences, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-8480, Japan.

出版信息

Mol Biol Rep. 2011 Jan;38(1):37-47. doi: 10.1007/s11033-010-0075-5. Epub 2010 Mar 20.

Abstract

Curved DNA structures with a left-handed superhelical conformation can activate eukaryotic transcription. However, their potency in transgene activation in embryonic stem (ES) cells has not been examined. T20 is an artificial curved DNA of 180 bp that serves as a transcriptional activator. We investigated the effect of T20 on transcription in mouse ES cell lines or hepatocytes differentiated from them. We established 10 sets of cell lines each harboring a single copy of the reporter construct. Each set comprised a T20-harboring cell line and a T20-less control cell line. Analyses showed that in ES cells and in hepatocytes originating from these cells, T20 both activated and repressed transcription in a manner that was dependent on the locus of reporter. The present and previous studies strongly suggest that in cells that have a strict gene regulation system, transcriptional activation by T20 occurs only in a transcriptionally active locus in the genome.

摘要

具有左手超螺旋构象的弯曲 DNA 结构可以激活真核转录。然而,它们在胚胎干细胞(ES)细胞中转基因激活的效力尚未被检测。T20 是一种 180bp 的人工弯曲 DNA,可作为转录激活剂。我们研究了 T20 对来自 ES 细胞系或由其分化而来的肝细胞中转录的影响。我们建立了 10 组细胞系,每组都含有一个报告基因构建体的单一拷贝。每组都包括一个携带 T20 的细胞系和一个不含 T20 的对照细胞系。分析表明,在 ES 细胞中和这些细胞来源的肝细胞中,T20 以依赖于报告基因座的方式激活和抑制转录。目前和以前的研究强烈表明,在具有严格基因调控系统的细胞中,T20 的转录激活仅发生在基因组中转录活跃的基因座上。

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