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Profiling of H3K4me3 Modification in Plants using Cleavage under Targets and Tagmentation

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Published on: April 22, 2022

Multi-CUT&Tag to simultaneously profile multiple chromatin factors.

Sneha Gopalan¹, Thomas G Fazzio¹

¹Department of Molecular, Cell, and Cancer Biology, University of Massachusetts Chan Medical School, Worcester, MA 01605, USA.

|January 31, 2022

Summary

We developed multi-CUT&Tag, a new method for simultaneously mapping multiple chromatin proteins in single cells. This technique allows for the direct detection of protein colocalization, advancing epigenomic profiling.

Keywords:

Antibody Genomics Molecular Biology Molecular/Chemical Probes Protein Biochemistry Protein expression and purification Sequence analysis Sequencing Single Cell

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Single-Cell Factor Localization on Chromatin using Ultra-Low Input Cleavage Under Targets and Release using Nuclease

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Published on: February 1, 2022

Area of Science:

Epigenetics
Molecular Biology
Genomics

Background:

Current genome-wide chromatin mapping methods typically analyze one protein at a time.
Simultaneous analysis of multiple chromatin proteins is crucial for understanding complex regulatory networks.

Purpose of the Study:

To introduce and validate multi-CUT&Tag, a novel technique for simultaneous mapping of multiple chromatin proteins.
To enable direct detection of protein colocalization within single cells or cell populations.

Main Methods:

Development of multi-CUT&Tag using barcoded adapters and antibody-protein A-Tn5 transposase complexes.
Application of the protocol for simultaneous chromatin protein mapping and colocalization analysis.

Main Results:

Multi-CUT&Tag successfully maps the locations of multiple chromatin proteins concurrently.
The method directly detects colocalization of different proteins within the same cell(s).

Conclusions:

Multi-CUT&Tag offers a powerful approach for generating multi-factor epigenomic maps.
This technique enhances the resolution of chromatin protein interaction studies compared to single-factor methods.