通过 RAT-ChIP 对牛囊胚的内细胞团和滋养外胚层进行全基因组组蛋白修饰谱分析。
Genome-wide histone modification profiling of inner cell mass and trophectoderm of bovine blastocysts by RAT-ChIP.
机构信息
Department of Biotechnology, Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.
Chair of Animal Breeding and Biotechnology, Estonian University of Life Sciences, Tartu, Estonia.
出版信息
PLoS One. 2019 Nov 25;14(11):e0225801. doi: 10.1371/journal.pone.0225801. eCollection 2019.
Chromatin immunoprecipitation coupled with next-generation sequencing (ChIP-seq) has revolutionized our understanding of chromatin-related biological processes. The method, however, requires thousands of cells and has therefore limited applications in situations where cell numbers are limited. Here we describe a novel method called Restriction Assisted Tagmentation Chromatin Immunoprecipitation (RAT-ChIP) that enables global histone modification profiling from as few as 100 cells. The method is simple, cost-effective and takes a single day to complete. We demonstrate the sensitivity of the method by deriving the first genome-wide maps of histone H3K4me3 and H3K27me3 modifications of inner cell mass and trophectoderm of bovine blastocyst stage embryos.
染色质免疫沉淀结合下一代测序（ChIP-seq）技术极大地改变了我们对与染色质相关的生物学过程的理解。然而，该方法需要数千个细胞，因此在细胞数量有限的情况下应用有限。在这里，我们描述了一种称为限制辅助标签化染色质免疫沉淀（RAT-ChIP）的新方法，该方法可从少至 100 个细胞中进行全局组蛋白修饰谱分析。该方法简单、经济高效，仅需一天即可完成。我们通过获得牛囊胚期胚胎内细胞团和滋养外胚层的组蛋白 H3K4me3 和 H3K27me3 修饰的全基因组图谱，证明了该方法的灵敏度。
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