使用超灵敏 STAR ChIP-Seq 技术在胚胎和低输入样本中追踪组蛋白修饰。
Tracking Histone Modifications in Embryos and Low-Input Samples Using Ultrasensitive STAR ChIP-Seq.
机构信息
Center for Stem Cell Biology and Regenerative Medicine, MOE Key Laboratory of Bioinformatics, THU-PKU Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing, China.
Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore.
出版信息
Methods Mol Biol. 2021;2214:241-252. doi: 10.1007/978-1-0716-0958-3_16.
ChIP-seq is a powerful technique that allows the detection of chromatin localization for proteins and epigenetic modifications. However, conventional ChIP-seq usually requires millions of cells. This becomes a daunting task for applications in which only limited experimental materials are available. For example, during mammalian embryo development, the epigenomes undergo drastic reprogramming which endows a fertilized egg with the potential to develop into the whole body. Low-input ChIP-seq methods would be instrumental to help decipher molecular mechanisms underlying such epigenetic reprogramming. Here we describe an optimized ChIP-seq method-STAR (Small-scale TELP-Assisted Rapid) ChIP-seq-that allows the detection of histone modifications using only a few hundred cells. This method is proven to be robust in epigenomic profiling in both embryos and cultured cells.
ChIP-seq 是一种强大的技术，可用于检测蛋白质和表观遗传修饰的染色质定位。然而，传统的 ChIP-seq 通常需要数百万个细胞。对于应用来说，这是一项艰巨的任务，因为实验材料有限。例如，在哺乳动物胚胎发育过程中，表观基因组经历了剧烈的重编程，赋予受精卵发育成整个身体的潜力。低输入 ChIP-seq 方法将有助于破译这种表观遗传重编程背后的分子机制。在这里，我们描述了一种优化的 ChIP-seq 方法——STAR（小规模 TELP 辅助快速）ChIP-seq——仅使用几百个细胞即可检测组蛋白修饰。该方法在胚胎和培养细胞的表观基因组分析中被证明是稳健的。
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