Cleavage Under Targets & Release Using Nuclease (CUT&RUN) is a groundbreaking approach for ultra-sensitive mapping of chromatin targets developed by the group of Dr. Steven Henikoff (see the paper). It builds on immunotethering technology (chromatin immunocleavage or ChIC) developed by Dr. Ulrich Laemmli (see this paper), wherein a fusion of Protein A to Micrococcal Nuclease (pA-MNase) is recruited to selectively cleave antibody-bound chromatin in intact cells or nuclei.

In CUTANA CUT&RUN, cells or nuclei are immobilized to a solid support and Protein A/Protein G-MNase (pAG-MNase) is used to selectively cleave antibody-labeled chromatin in intact cells (see Figure 1 for workflow overview). The clipped fragments diffuse into solution, where they can be separated from cells, purified, and analyzed by next-generation sequencing. This workflow results in high-quality, genome-wide profiles of histone post-translational modifications (PTMs) and chromatin-associated proteins (e.g. transcription factors).

Figure 1. Overview of CUTANA CUT&RUN workflow.

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