表观遗传学
转座酶
计算生物学
染色质
组蛋白
生物
DNA甲基化
分子生物学
细胞生物学
基因表达
基因
遗传学
基因组
转座因子
作者
Hatice S Kaya-Okur,Steven J. Wu,Christine A. Codomo,Erica S. Pledger,Terri D. Bryson,Steven Henikoff,Kami Ahmad,Steven Henikoff
摘要
Abstract Many chromatin features play critical roles in regulating gene expression. A complete understanding of gene regulation will require the mapping of specific chromatin features in small samples of cells at high resolution. Here we describe Cleavage Under Targets and Tagmentation (CUT&Tag), an enzyme-tethering strategy that provides efficient high-resolution sequencing libraries for profiling diverse chromatin components. In CUT&Tag, a chromatin protein is bound in situ by a specific antibody, which then tethers a protein A-Tn5 transposase fusion protein. Activation of the transposase efficiently generates fragment libraries with high resolution and exceptionally low background. All steps from live cells to sequencing-ready libraries can be performed in a single tube on the benchtop or a microwell in a high-throughput pipeline, and the entire procedure can be performed in one day. We demonstrate the utility of CUT&Tag by profiling histone modifications, RNA Polymerase II and transcription factors on low cell numbers and single cells.
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