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CUT&Tag for efficient epigenomic profiling of small samples and single cells

Hatice S. Kaya-Okur, Steven J. Wu, Christine A. Codomo, Erica S. Pledger, Terri D. Bryson, View ORCID ProfileJorja G. Henikoff, Kami Ahmad, Steven Henikoff
doi: https://doi.org/10.1101/568915
Hatice S. Kaya-Okur
1Basic Sciences Division, Fred Hutchinson Cancer Research Center, 1100 N. Fairview Ave, Seattle, WA, 98109
2Howard Hughes Medical Institute, USA
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Steven J. Wu
1Basic Sciences Division, Fred Hutchinson Cancer Research Center, 1100 N. Fairview Ave, Seattle, WA, 98109
3Molecular Engineering & Sciences Institute, University of Washington, Seattle, WA, 98195, USA
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Christine A. Codomo
1Basic Sciences Division, Fred Hutchinson Cancer Research Center, 1100 N. Fairview Ave, Seattle, WA, 98109
2Howard Hughes Medical Institute, USA
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Erica S. Pledger
1Basic Sciences Division, Fred Hutchinson Cancer Research Center, 1100 N. Fairview Ave, Seattle, WA, 98109
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Terri D. Bryson
1Basic Sciences Division, Fred Hutchinson Cancer Research Center, 1100 N. Fairview Ave, Seattle, WA, 98109
2Howard Hughes Medical Institute, USA
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Jorja G. Henikoff
1Basic Sciences Division, Fred Hutchinson Cancer Research Center, 1100 N. Fairview Ave, Seattle, WA, 98109
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  • ORCID record for Jorja G. Henikoff
Kami Ahmad
1Basic Sciences Division, Fred Hutchinson Cancer Research Center, 1100 N. Fairview Ave, Seattle, WA, 98109
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Steven Henikoff
1Basic Sciences Division, Fred Hutchinson Cancer Research Center, 1100 N. Fairview Ave, Seattle, WA, 98109
2Howard Hughes Medical Institute, USA
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Abstract

Many chromatin features play critical roles in regulating gene expression. A complete understanding of gene regulation will require the mapping of specific chromatin features in small samples of cells at high resolution. Here we describe Cleavage Under Targets and Tagmentation (CUT&Tag), an enzyme-tethering strategy that provides efficient high-resolution sequencing libraries for profiling diverse chromatin components. In CUT&Tag, a chromatin protein is bound in situ by a specific antibody, which then tethers a protein A-Tn5 transposase fusion protein. Activation of the transposase efficiently generates fragment libraries with high resolution and exceptionally low background. All steps from live cells to sequencing-ready libraries can be performed in a single tube on the benchtop or a microwell in a high-throughput pipeline, and the entire procedure can be performed in one day. We demonstrate the utility of CUT&Tag by profiling histone modifications, RNA Polymerase II and transcription factors on low cell numbers and single cells.

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Posted March 06, 2019.
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CUT&Tag for efficient epigenomic profiling of small samples and single cells
Hatice S. Kaya-Okur, Steven J. Wu, Christine A. Codomo, Erica S. Pledger, Terri D. Bryson, Jorja G. Henikoff, Kami Ahmad, Steven Henikoff
bioRxiv 568915; doi: https://doi.org/10.1101/568915
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CUT&Tag for efficient epigenomic profiling of small samples and single cells
Hatice S. Kaya-Okur, Steven J. Wu, Christine A. Codomo, Erica S. Pledger, Terri D. Bryson, Jorja G. Henikoff, Kami Ahmad, Steven Henikoff
bioRxiv 568915; doi: https://doi.org/10.1101/568915

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