Journal Article

CUT&Tag for efficient epigenomic profiling of small samples and single cells

Henikoff, Jorja G., Pledger, Erica S., Wu, Steven J., Ahmad, Kami, Henikoff, Steven, Codomo, Christine A., Kaya-Okur, Hatice S., Bryson, Terri D.

2019

Overview

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. Understanding gene regulation will require mapping specific chromain features in a small number of cells at high resolution. Here the authors describe CUT&Tag, which uses antibody-mediated tethering of Tn5 transposase to a chromatin protein to generate high resolution libraries.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 45 / 45/15 / 631/1647 / 631/1647/2210 / 631/1647/2210/2211 / 631/61 / 631/61/212 / Chromatin / Chromatin - chemistry / Chromatin - metabolism / DNA-directed RNA polymerase / Epigenomics - methods / Fusion protein / Gene expression / Gene Expression Profiling - methods / Gene Expression Regulation / Gene mapping / Gene regulation / Genomic Library / High resolution / High-Throughput Nucleotide Sequencing / Histone Code / Histones - genetics / Histones - metabolism / Humanities and Social Sciences / Humans / K562 Cells / Libraries / Mapping / multidisciplinary / Protein A / Proteins / Recombinant Fusion Proteins - genetics / Recombinant Fusion Proteins - metabolism / Ribonucleic acid / RNA / RNA polymerase / RNA polymerase II / RNA Polymerase II - genetics / RNA Polymerase II - metabolism / Science / Science (multidisciplinary) / Single-Cell Analysis - methods / Staining and Labeling - methods / Staphylococcal Protein A - genetics / Staphylococcal Protein A - metabolism / Tethering / Tethers / Transcription factors / Transcription Factors - genetics / Transcription Factors - metabolism / Transposase / Transposases - genetics / Transposases - metabolism