PAtCh-Cap: input strategy for improving analysis of ChIP-exo data sets and beyond.

PAtCh-Cap: input strategy for improving analysis of ChIP-exo data sets and beyond.
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DOI:
10.1093/nar/gkw741
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发表时间:
2016-12-01
影响因子:
14.9
通讯作者:
Buck-Koehntop BA
Buck-Koehntop BA
中科院分区:
生物学2区
文献类型:
--
作者:
Terooatea TW;Pozner A;Buck-Koehntop BA

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最近,核心 ChIP-seq(染色质免疫沉淀结合下一代测序)方法取得了多项进展,以简化流程、降低成本或提高数据分辨率。其中一些新兴的基于 ChIP 的方法对珠子结合的免疫沉淀染色质执行额外的化学步骤,这对生成生物信息学分析期间去除伪影所需的类似处理的输入对照提出了挑战。在这里,我们提出了一种通用方法,用于为基于 ChIP 的方法产生技术特定的输入控制,该方法利用额外的珠结合处理步骤。这种报道的方法被称为蛋白质附着染色质捕获 (PAtCh-Cap),依赖于通过其羧基对染色质结合蛋白进行非特异性捕获,使 DNA 可用于后续化学处理,同时对目标蛋白进行染色质单独免疫沉淀。这种输入策略的应用不仅显着增强了 ChIP-exo 数据中的伪影去除,增加了峰识别的置信度并允许从头进行基序搜索,而且还发现了一种新的 CTCF 结合基序。
Recently, a number of advances have been implemented into the core ChIP-seq (chromatin immunoprecipitation coupled with next-generation sequencing) methodology to streamline the process, reduce costs or improve data resolution. Several of these emerging ChIP-based methods perform additional chemical steps on bead-bound immunoprecipitated chromatin, posing a challenge for generating similarly treated input controls required for artifact removal during bioinformatics analyses. Here we present a versatile method for producing technique-specific input controls for ChIP-based methods that utilize additional bead-bound processing steps. This reported method, termed protein attached chromatin capture (PAtCh-Cap), relies on the non-specific capture of chromatin-bound proteins via their carboxylate groups, leaving the DNA accessible for subsequent chemical treatments in parallel with chromatin separately immunoprecipitated for the target protein. Application of this input strategy not only significantly enhanced artifact removal from ChIP-exo data, increasing confidence in peak identification and allowing for de novo motif searching, but also afforded discovery of a novel CTCF binding motif.
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