Isolation of Specific Genomic Regions and Identification of Their Associated Molecules by Engineered DNA-Binding Molecule-Mediated Chromatin Immunoprecipitation (enChIP) Using the CRISPR System and TAL Proteins.
Isolation of Specific Genomic Regions and Identification of Their Associated Molecules by Engineered DNA-Binding Molecule-Mediated Chromatin Immunoprecipitation (enChIP) Using the CRISPR System and TAL Proteins.
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DOI：
10.3390/ijms160921802
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发表时间：
2015-09-09
影响因子：
5.6
通讯作者：
Fujita T
中科院分区：
文献类型：
--
作者：
Fujii H;Fujita T
全面了解基因组功能需要识别分子（蛋白质，RNA，基因组区域等）。与体内感兴趣的特定基因组区域结合。为了对特定基因组区域进行生物化学和分子生物学分析，我们开发了工程化DNA结合分子介导的染色质免疫沉淀（enChIP）来纯化感兴趣的基因组区域。在enChIP中，特定的基因组区域被标记用于使用工程化的DNA结合分子进行生物化学纯化，例如转录激活因子样（TAL）蛋白和成簇的规则间隔短回文重复序列（CRISPR）系统的催化失活形式。enChIP是一种综合方法，强调使用下一代测序（NGS），微阵列，质谱（MS）和其他方法进行无偏倚搜索。此外，这种方法不限于培养的细胞系，并且可以容易地扩展到生物体。在这篇综述中，我们将讨论enChIP的应用，阐明基因组功能的分子机制。
Comprehensive understanding of genome functions requires identification of molecules (proteins, RNAs, genomic regions, etc.) bound to specific genomic regions of interest in vivo. To perform biochemical and molecular biological analysis of specific genomic regions, we developed engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) to purify genomic regions of interest. In enChIP, specific genomic regions are tagged for biochemical purification using engineered DNA-binding molecules, such as transcription activator-like (TAL) proteins and a catalytically inactive form of the clustered regularly interspaced short palindromic repeats (CRISPR) system. enChIP is a comprehensive approach that emphasizes non-biased search using next-generation sequencing (NGS), microarrays, mass spectrometry (MS), and other methods. Moreover, this approach is not restricted to cultured cell lines and can be easily extended to organisms. In this review, we discuss applications of enChIP to elucidating the molecular mechanisms underlying genome functions.