1. Chromosomes and Gene Expression
Download icon

Chromatin Mapping: A cut above

  1. Chongsheng He
  2. Roberto Bonasio  Is a corresponding author
  1. Perelman School of Medicine, University of Pennsylvania, United States
Insight
Cite this article as: eLife 2017;6:e25000 doi: 10.7554/eLife.25000
1 figure

Figures

X-ChIP, native ChIP, and CUT&RUN.

(A) In X-ChIP, cells are first crosslinked (red crosses) with formaldehyde to freeze the interactions between the DNA (black line) and a chromatin-binding protein of interest (CP; blue). Sonication fragments the chromatin and makes it soluble. Antibodies are used to recognize the protein–DNA fragments, which are then ‘pulled’ out of the solution using antibody-binding beads, in a process called immunoprecipitation. The histones are shown in yellow. (B) In native ChIP, chromatin is fragmented and solubilized by treating cells with an enzyme called micrococcal nuclease (MNase; small brown shapes). The natural affinity of the protein for its DNA target keep them together during the immunoprecipitation process. (C) In CUT&RUN, antibodies direct the activity of the MNase enzyme to ensure that chromatin cleavage happens close to the protein of interest. A protein called protein A (brown ovals) binds the MNase enzyme to the antibody. The resulting small DNA fragments can be isolated as they diffuse out of the nuclei.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)