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CUT&Tag for efficient epigenomic profiling of small samples and single cells
Hatice S Kaya-Okur1,2, Steven J Wu1,3, Christine A Codomo1,2
1Basic Sciences Division, Fred Hutchinson Cancer Research Center, 1100N. FairviewAve, Seattle, WA, 98109, USA.
Nature Communications
|May 1, 2019
Summary
Cleavage Under Targets and Tagmentation (CUT&Tag) enables high-resolution mapping of chromatin features. This efficient method allows for rapid profiling of gene regulation components in single cells.
Area of Science:
- Molecular Biology
- Genomics
- Epigenetics
Background:
- Chromatin features are crucial for gene expression regulation.
- High-resolution mapping of these features in small cell samples is needed for a complete understanding of gene regulation.
Purpose of the Study:
- To introduce Cleavage Under Targets and Tagmentation (CUT&Tag), a novel enzyme-tethering strategy.
- To demonstrate the utility of CUT&Tag for profiling diverse chromatin components at high resolution.
Main Methods:
- CUT&Tag utilizes an antibody to bind a target chromatin protein in situ.
- A Protein A-Tn5 transposase fusion protein is tethered to the antibody.
- Transposase activation generates high-resolution sequencing libraries with low background.
Main Results:
- The CUT&Tag protocol allows for efficient library generation from live cells to sequencing-ready libraries in a single tube.
- The entire procedure can be completed in one day.
- CUT&Tag successfully profiled histone modifications, RNA Polymerase II, and transcription factors in low cell numbers and single cells.
Conclusions:
- CUT&Tag is an efficient and high-resolution method for profiling chromatin components.
- This technique facilitates the study of gene regulation in limited cell populations and single cells.
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