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采用染色质免疫沉淀和大规模平行测序技术对红细胞白血病细胞内 EGR1 结合位点进行体内的全基因组分析。

Global analysis of in vivo EGR1-binding sites in erythroleukemia cell using chromatin immunoprecipitation and massively parallel sequencing.

机构信息

School of Basic Medical Sciences, Southeast University, Nanjing, P. R. China.

出版信息

Electrophoresis. 2010 Sep;31(17):2936-43. doi: 10.1002/elps.201000094.

Abstract

Early growth response gene 1 (EGR1) has been implicated in megakaryocyte differentiation induced by phorbol ester. But the molecule mechanism of EGR1 in this process has not been widely investigated. The identification of direct EGR1 target genes in a global scale is critical for our understanding of how EGR1 contributes to this process. In this study, we provide a global survey on the binding location of EGR1 in the K562 cells using chromatin immunoprecipitation and massively parallel sequencing. Over 14 000 highly confident in vivo EGR1 binding sites were identified in phorbol 12-myristate 13-acetate-treated K562 cells. More than 70% of these genomic sites associated with EGR1 binding were located around annotated gene regions. Molecular functional classification of 6138 putative EGR1 target genes showed that the transcription factor class (695 of 6138; 11%) is the largest significantly enriched one. The results showed that a high coverage of the genome and a high positive rate achieve were achieved. This whole genome study on the EGR1 targets may provide a better understanding of the EGR1 regulated genes and the downstream pathway in megakaryocyte differentiation.

摘要

早期生长反应基因 1(EGR1)已被牵涉到佛波酯诱导的巨核细胞分化中。但是,EGR1 在这个过程中的分子机制还没有被广泛研究。在全局范围内鉴定直接的 EGR1 靶基因对于我们理解 EGR1 如何促进这个过程至关重要。在这项研究中,我们使用染色质免疫沉淀和大规模平行测序,对 PMA 处理的 K562 细胞中 EGR1 的结合位置进行了全面调查。在 PMA 处理的 K562 细胞中,鉴定出了超过 14000 个高度可信的体内 EGR1 结合位点。这些与 EGR1 结合相关的基因组位点中,超过 70%位于注释基因区域附近。6138 个假定的 EGR1 靶基因的分子功能分类表明,转录因子类(6138 个中的 695 个;11%)是最大的显著富集类。结果表明,实现了基因组的高覆盖率和高阳性率。这项关于 EGR1 靶标的全基因组研究可能为理解 EGR1 调节的基因和巨核细胞分化的下游途径提供更好的认识。

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