人类原始多能干细胞的染色质分析。

Chromatin Profiling of Human Naïve Pluripotent Stem Cells.

机构信息

Epigenetics Programme, The Babraham Institute, Cambridge, UK.

出版信息

Methods Mol Biol. 2022;2416:181-200. doi: 10.1007/978-1-0716-1908-7_12.

DOI:10.1007/978-1-0716-1908-7_12

Abstract

Chromatin immunoprecipitation combined with high-throughput sequencing (ChIP-sequencing) facilitates the genome-wide mapping of DNA sequences that are enriched for specific chromatin-binding proteins or histone post-translational modifications. More recently developed chromatin profiling methods called Cleavage Under Targets and Release Using Nuclease (CUT&RUN) and Cleavage Under Targets and Tagmentation (CUT&Tag) have adapted the ChIP-sequencing approach to produce similar data from a smaller amount of starting material, and while overcoming many of the conventional drawbacks of ChIP-sequencing. Here, we present detailed protocols for ChIP-seq, CUT&RUN, and CUT&Tag to profile genome-wide protein-DNA interactions in naïve human pluripotent stem cells.

摘要

染色质免疫沉淀结合高通量测序（ChIP-seq）可促进对特定染色质结合蛋白或组蛋白翻译后修饰富集的 DNA 序列进行全基因组作图。最近开发的染色质分析方法，如靶向切割和释放使用核酸酶（CUT&RUN）和靶向切割和标签化（CUT&Tag），已经对 ChIP-seq 方法进行了改进，可从更少的起始材料中产生类似的数据，并克服了 ChIP-seq 的许多传统缺点。在这里，我们提供了用于 ChIP-seq、CUT&RUN 和 CUT&Tag 的详细方案，以分析原始人多能干细胞中的全基因组蛋白-DNA 相互作用。