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Improved CUT&RUN chromatin profiling tools
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Journal Article
Improved CUT&RUN chromatin profiling tools
2019
Overview
Previously, we described a novel alternative to chromatin immunoprecipitation, CUT&RUN, in which unfixed permeabilized cells are incubated with antibody, followed by binding of a protein A-Micrococcal Nuclease (pA/MNase) fusion protein (Skene and Henikoff, 2017). Here we introduce three enhancements to CUT&RUN: A hybrid protein A-Protein G-MNase construct that expands antibody compatibility and simplifies purification, a modified digestion protocol that inhibits premature release of the nuclease-bound complex, and a calibration strategy based on carry-over of E. coli DNA introduced with the fusion protein. These new features, coupled with the previously described low-cost, high efficiency, high reproducibility and high-throughput capability of CUT&RUN make it the method of choice for routine epigenomic profiling.
Publisher
eLife Science Publications, Ltd,eLife Sciences Publications Ltd,eLife Sciences Publications, Ltd
Subject
/ Bacterial Proteins - genetics
/ Bacterial Proteins - metabolism
/ Chromosomes and Gene Expression
/ DNA
/ Genomes
/ Immunologic Techniques - methods
/ Methods
/ Micrococcal Nuclease - genetics
/ Micrococcal Nuclease - metabolism
/ Novels
/ Nuclease
/ Proteins
/ Protocol
/ Recombinant Fusion Proteins - genetics
/ Recombinant Fusion Proteins - metabolism
/ Staphylococcal Protein A - genetics
