利用基因座特异性染色质免疫沉淀技术对基因组功能进行生化分析

T. Fujita, H. Fujii
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引用次数: 12

摘要

为了分离保留其分子相互作用的特定基因组区域,允许直接鉴定染色质结合分子,我们开发了两种基因座特异性染色质免疫沉淀(基因座特异性ChIP)技术,插入芯片(iChIP)和工程dna结合分子介导的芯片(enChIP),使用集群规则间隔短回文重复序列(CRISPR)系统或转录激活因子样(TAL)蛋白。从本质上讲,基因座特异性ChIP包括基因座标记和亲和纯化,并可与下游分析相结合,以识别与目标基因组区域相关的分子。在这篇综述中,我们讨论了基因座特异性芯片在基因组功能分析中的应用,包括转录和表观遗传调控。
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Biochemical Analysis of Genome Functions Using Locus-Specific Chromatin Immunoprecipitation Technologies
To isolate specific genomic regions that retain their molecular interactions, allowing direct identification of chromatin-bound molecules, we developed two locus-specific chromatin immunoprecipitation (locus-specific ChIP) technologies, insertional ChIP (iChIP) and engineered DNA-binding molecule-mediated ChIP (enChIP) using the clustered regularly interspaced short palindromic repeats (CRISPR) system or transcription activator-like (TAL) proteins. Essentially, a locus-specific ChIP consists of locus-tagging and affinity purification and can be combined with downstream analyses to identify molecules associated with the target genomic regions. In this review, we discuss the applications of locus-specific ChIP to analyze the genome functions, including transcription and epigenetic regulation.
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