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通过染色质免疫沉淀展示技术鉴定转录因子靶基因

Identification of transcription factor target genes by ChIP display.

作者信息

Barski Artem, Pregizer Steven, Frenkel Baruch

机构信息

Department of Biochemistry & Molecular Biology, University of Southern California Keck School of Medicine, Los Angeles, CA, USA.

出版信息

Methods Mol Biol. 2008;455:177-90. doi: 10.1007/978-1-59745-104-8_14.

Abstract

Transcription factors play pivotal roles in the control of cell growth and differentiation in health and disease. In the post-genomic era, it has become possible to locate the regions occupied by transcription factors throughout the genome, leading to better understanding of their mechanism of action and the genes that they regulate. All methods for transcription factor location analysis utilize chromatin immunoprecipitation (ChIP). Although ChIP was initially used to test whether a protein binds to a candidate promoter in living cells, newly developed methods allow the unbiased identification of novel targets of transcription factors. This chapter describes ChIP Display, an affordable method for transcription factor location analysis. Despite being relatively low throughput compared with alternative methods such as ChIP-chip and ChIP-SAGE, ChIP Display provides even small molecular biology laboratories with the opportunity to discover novel targets of any transcription factor, for which high-quality antibodies are available.

摘要

转录因子在健康和疾病状态下细胞生长与分化的调控中发挥着关键作用。在后基因组时代,已能够定位转录因子在整个基因组中占据的区域,从而更深入地了解其作用机制以及它们所调控的基因。所有用于转录因子定位分析的方法都利用了染色质免疫沉淀技术(ChIP)。尽管ChIP最初用于检测蛋白质是否在活细胞中与候选启动子结合,但新开发的方法能够无偏向性地鉴定转录因子的新靶点。本章介绍ChIP Display,这是一种用于转录因子定位分析的经济实惠的方法。尽管与ChIP-chip和ChIP-SAGE等其他方法相比,其通量相对较低,但ChIP Display为即使是小型分子生物学实验室提供了发现任何转录因子新靶点的机会,前提是有针对该转录因子的高质量抗体。

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