Efficient low-cost chromatin profiling with CUT&Tag

By Hatice S. Kaya-Okur, Derek H. Janssens, Jorja G. Henikoff, K. Ahmad, Steven Henikoff. Nature Protocols, 2020

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We recently introduced Cleavage Under Targets & Tagmentation (CUT&Tag), an epigenomic profiling strategy in which antibodies are bound to chromatin proteins in situ in permeabilized nuclei. These antibodies are then used to tether the cut-and-paste transposase Tn5. Activation of the transposase simultaneously cleaves DNA and adds adapters (‘tagmentation’) for paired-end DNA sequencing. Here, we introduce a streamlined CUT&Tag protocol that suppresses DNA accessibility artefacts to ensure high-fidelity mapping of the antibody-targeted protein and improves the signal-to-noise ratio over current chromatin profiling methods. Streamlined CUT&Tag can be performed in a single PCR tube, from cells to amplified libraries, providing low-cost genome-wide chromatin maps. By simplifying library preparation CUT&Tag requires less than a day at the bench, from live cells to sequencing-ready barcoded libraries. As a result of low background levels, barcoded and pooled CUT&Tag libraries can be sequenced for as little as $25 per sample. This enables routine genome-wide profiling of chromatin proteins and modifications and requires no special skills or equipment. The authors describe a streamlined epigenomic profiling protocol based on cut-and-paste tagmentation by the Tn5 transposase targeted to a chromatin protein of interest.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/66817dcd5ccf1c8cd14125346d5be1e297959505