Improved CUT&RUN chromatin profiling tools.
Michael P MeersTerri D BrysonJorja G HenikoffSteven HenikoffPublished in: eLife (2019)
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.
Keyphrases
- low cost
- high efficiency
- high throughput
- protein protein
- gene expression
- single cell
- transcription factor
- dna damage
- binding protein
- induced apoptosis
- randomized controlled trial
- dna binding
- genome wide
- amino acid
- escherichia coli
- small molecule
- cell cycle arrest
- cell free
- cell death
- signaling pathway
- dna methylation
- cell proliferation
- single molecule
- endoplasmic reticulum stress
- decision making
- recombinant human