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Improved CUT&RUN chromatin profiling tools.
Elife 2019; 8E

Abstract

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.

Authors+Show Affiliations

Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, United States.Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, United States. Howard Hughes Medical Institute, United States.Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, United States.Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, United States. Howard Hughes Medical Institute, United States.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

31232687

Citation

Meers, Michael P., et al. "Improved CUT&RUN Chromatin Profiling Tools." ELife, vol. 8, 2019.
Meers MP, Bryson TD, Henikoff JG, et al. Improved CUT&RUN chromatin profiling tools. Elife. 2019;8.
Meers, M. P., Bryson, T. D., Henikoff, J. G., & Henikoff, S. (2019). Improved CUT&RUN chromatin profiling tools. ELife, 8, doi:10.7554/eLife.46314.
Meers MP, et al. Improved CUT&RUN Chromatin Profiling Tools. Elife. 2019 06 24;8 PubMed PMID: 31232687.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Improved CUT&RUN chromatin profiling tools. AU - Meers,Michael P, AU - Bryson,Terri D, AU - Henikoff,Jorja G, AU - Henikoff,Steven, Y1 - 2019/06/24/ PY - 2019/02/22/received PY - 2019/06/22/accepted PY - 2019/6/25/pubmed PY - 2019/6/25/medline PY - 2019/6/25/entrez KW - chromatin KW - chromosomes KW - epigenomics KW - gene expression KW - genetics KW - genomics KW - human KW - spike-in calibration JF - eLife JO - Elife VL - 8 N2 - 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. SN - 2050-084X UR - https://www.unboundmedicine.com/medline/citation/31232687/Improved_CUTRUN_chromatin_profiling_tools L2 - https://doi.org/10.7554/eLife.46314 DB - PRIME DP - Unbound Medicine ER -