1. CUT&Tag for efficient epigenomic profiling of small samples and single cells.
- Author
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Kaya-Okur HS, Wu SJ, Codomo CA, Pledger ES, Bryson TD, Henikoff JG, Ahmad K, and Henikoff S
- Subjects
- Chromatin metabolism, Gene Expression Regulation, Genomic Library, High-Throughput Nucleotide Sequencing, Histone Code, Histones genetics, Histones metabolism, Humans, K562 Cells, RNA Polymerase II genetics, RNA Polymerase II metabolism, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Staphylococcal Protein A genetics, Staphylococcal Protein A metabolism, Transcription Factors genetics, Transcription Factors metabolism, Transposases genetics, Transposases metabolism, Chromatin chemistry, Epigenomics methods, Gene Expression Profiling methods, Single-Cell Analysis methods, Staining and Labeling methods
- 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.
- Published
- 2019
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