MicroChIP--a rapid micro chromatin immunoprecipitation assay for small cell samples and biopsies.
MicroChIP--a rapid micro chromatin immunoprecipitation assay for small cell samples and biopsies.
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DOI：
10.1093/nar/gkm1158
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发表时间：
2008-02
影响因子：
14.9
通讯作者：
Collas P
中科院分区：
文献类型：
--
作者：
Dahl JA;Collas P
染色质免疫沉淀（ChIP）是研究蛋白质-DNA相互作用的有力技术。然而，目前ChIP测定的缺点是需要大量细胞，这限制了ChIP对稀有细胞样品的适用性，和/或限制了应用的冗长程序。迄今为止，还没有用于来自小细胞数目或活检的后修饰组蛋白的快速和平行ChIP的方案，并且重要的是，没有允许研究小细胞数目中的转录因子结合的方案。我们在此报告了一种微（μ）ChIP检测方法的开发，该方法适用于单批1000个细胞中修饰组蛋白或RNA聚合酶II的多达9个平行定量ChIP。μChIP也可以缩小规模，以监测一种蛋白质与100个细胞中多个基因组位点的关联。μChIP适用于小的新鲜组织活检，解冻时交联程序使该测定法适用于冷冻活检。使用μChIP，我们表征了人类胚胎癌细胞和骨肉瘤活检组织中发育调控启动子上的转录允许和抑制性组蛋白H3修饰。μChIP为稀有细胞和组织样本的多重平行和快速转录因子结合和表观遗传分析创造了可能性。
Chromatin immunoprecipitation (ChIP) is a powerful technique for studying protein–DNA interactions. Drawbacks of current ChIP assays however are a requirement for large cell numbers, which limits applicability of ChIP to rare cell samples, and/or lengthy procedures with limited applications. There are to date no protocols for fast and parallel ChIPs of post-translationally modified histones from small cell numbers or biopsies, and importantly, no protocol allowing for investigations of transcription factor binding in small cell numbers. We report here the development of a micro (μ) ChIP assay suitable for up to nine parallel quantitative ChIPs of modified histones or RNA polymerase II from a single batch of 1000 cells. μChIP can also be downscaled to monitor the association of one protein with multiple genomic sites in as few as 100 cells. μChIP is applicable to small fresh tissue biopsies, and a cross-link-while-thawing procedure makes the assay suitable for frozen biopsies. Using μChIP, we characterize transcriptionally permissive and repressive histone H3 modifications on developmentally regulated promoters in human embryonal carcinoma cells and in osteosarcoma biopsies. μChIP creates possibilities for multiple parallel and rapid transcription factor binding and epigenetic analyses of rare cell and tissue samples.
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DOI：
10.1073/pnas.0510419103
发表时间：
2006-01-31
影响因子：
11.1
作者：
Ge, WH;He, F;Sun, YE
通讯作者：
Sun, YE
影响因子：
64.5
作者：
Guenther, Matthew G.;Levine, Stuart S.;Young, Richard A.
通讯作者：
Young, Richard A.
影响因子：
5.3
作者：
Zuccato, Chiara;Belyaev, Nikolai;Cattaneo, Elena
通讯作者：
Cattaneo, Elena
影响因子：
64.8
作者：
SCHOLER, HR;RUPPERT, S;GRUSS, P
通讯作者：
GRUSS, P
快速染色质免疫沉淀测定法。
影响因子：
14.9
作者：
通讯作者：
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