Paper

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

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

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.

• Publisher: Cold Spring Harbor Laboratory Press,Cold Spring Harbor Laboratory
• Subject: Chromatin / DNA-directed RNA polymerase / Fusion protein / Gene expression / Gene mapping / Gene regulation / Genomics / Libraries / Proteins / RNA polymerase / Transcription factors / Transposase