Fujita Toshitsugu, Fujii Hodaka
Combined Program on Microbiology and Immunology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
ISRN Biochem. 2013 Jan 10;2013:913273. doi: 10.1155/2013/913273. eCollection 2013.
Comprehensive understanding of regulation mechanisms of biological phenomena mediated by functions of genomic DNA requires identification of molecules bound to genomic regions of interest in vivo. However, nonbiased methods to identify molecules bound to specific genomic loci in vivo are limited. To perform biochemical and molecular biological analysis of specific genomic regions, we developed the insertional chromatin immunoprecipitation (iChIP) technology to purify the genomic regions of interest. We applied iChIP to direct identification of components of insulator complexes, which function as boundaries of chromatin domain, showing that it is feasible to directly identify proteins and RNA bound to a specific genomic region in vivo by using iChIP. In addition, recently, we succeeded in identifying proteins and genomic regions interacting with a single copy endogenous locus. In this paper, we will discuss the application of iChIP to epigenetics and chromatin research.
全面了解由基因组DNA功能介导的生物现象的调控机制需要鉴定体内与感兴趣的基因组区域结合的分子。然而，在体内鉴定与特定基因组位点结合的分子的无偏方法有限。为了对特定基因组区域进行生化和分子生物学分析，我们开发了插入染色质免疫沉淀（iChIP）技术来纯化感兴趣的基因组区域。我们将iChIP应用于直接鉴定绝缘子复合物的成分，该复合物作为染色质结构域的边界，表明使用iChIP在体内直接鉴定与特定基因组区域结合的蛋白质和RNA是可行的。此外，最近我们成功鉴定了与单拷贝内源性位点相互作用的蛋白质和基因组区域。在本文中，我们将讨论iChIP在表观遗传学和染色质研究中的应用。
