released
December 13th, 2019 at 6:53pm
Overview
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Assay Description
pA-DamID
The proteinA-DamID (pA-DamID) method is a hybrid of the CUT&RUN (Skene, 2017) and DamID (Vogel, 2007) technologies.
Cells are permeabilized with digitonin and incubated with an antibody against a nuclear protein of interest, followed by binding with pA-Dam. Subsequent activation of the tethered Dam by its methyldonor S-adenosyl-methionine (SAM) results in in situ Gm6ATC methylation of DNA sequences that are in molecular proximity to the protein of interest. The pattern of deposited m6A can be mapped genome-wide as in conventional DamID. The labeled DNA can also be visualized using the m6A-Tracer (Kind, 2013), providing a powerful quality control and possibly new biological insights. In addition, labeled cells can be sorted to study (rare) subpopulations of interest.
Image source: van Steensel Lab