Journal Article

Targeted in situ genome-wide profiling with high efficiency for low cell numbers

Skene, Peter J, Henikoff, Steven, Henikoff, Jorja G

2018

Overview

Cleavage under targets and release using nuclease (CUT&RUN) is an epigenomic profiling strategy in which antibody-targeted controlled cleavage by micrococcal nuclease releases specific protein-DNA complexes into the supernatant for paired-end DNA sequencing. As only the targeted fragments enter into solution, and the vast majority of DNA is left behind, CUT&RUN has exceptionally low background levels. CUT&RUN outperforms the most widely used chromatin immunoprecipitation (ChIP) protocols in resolution, signal-to-noise ratio and depth of sequencing required. In contrast to ChIP, CUT&RUN is free of solubility and DNA accessibility artifacts and has been used to profile insoluble chromatin and to detect long-range 3D contacts without cross-linking. Here, we present an improved CUT&RUN protocol that does not require isolation of nuclei and provides high-quality data when starting with only 100 cells for a histone modification and 1,000 cells for a transcription factor. From cells to purified DNA, CUT&RUN requires less than a day at the laboratory bench and requires no specialized skills.

• Publisher: Nature Publishing Group
• Subject: Background levels / Chromatin / Cleavage / Crosslinking / Deoxyribonucleic acid / DNA / DNA sequencing / Genomes / Immunoprecipitation / Nuclease / Nuclei / Nuclei (cytology)