Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq)
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Abstract
Modern next-generation sequencing-based methods have empowered researchers to assay the epigenetic states of individual cells. Existing techniques for profiling epigenetic marks in single cells often require the use and optimization of time-intensive procedures such as drop fluidics, chromatin fragmentation, and end repair. Here we describe ACT-seq, a streamlined method for mapping genome-wide distributions of histone tail modifications, histone variants, and chromatin-binding proteins in a small number of or single cells. ACT-seq utilizes a fusion of Tn5 transposase to Protein A that is targeted to chromatin by a specific antibody, allowing chromatin fragmentation and sequence tag insertion specifically at genomic sites presenting the relevant antigen. The Tn5 transposase enables the use of an index multiplexing strategy (iACT-seq), which enables construction of thousands of single-cell libraries in one day by a single researcher without the need for drop-based fluidics or visual sorting. We conclude that ACT-seq present an attractive alternative to existing techniques for mapping epigenetic marks in single cells.
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Single-cell characterization of histone post-translational modifications has not been achieved for highly complex adult organs such as the brain.
“However, for highly complex adult organs such as the brain, this single-cell characterization has not been achieved.”
From Single-cell CUT&Tag profiles histone modifications and transcription factors in complex tissues (2021) · cites this paper as [11]
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- Authors
- Carter, B., Ku, W. L., Kang, J. Y., Hu, G., Perrie, J., Tang, Q., & Zhao, K.
- Affiliations
- National Institutes of Health (US)
- Corresponding author
- Zhao, Keji
- Venue
- Nature Communications
- Published
- 2019-08-20
- Volume
- 10
- Issue
- 1
- Pages
- 3747-3747
- ISSN
- 2041-1723 (ISSN-L)
- PMC
- PMC6702168
- OpenAlex
- W2969437743
- Cited by
- 148
- References
- 22
- Citations by year
- 2026: 8 · 2025: 16 · 2024: 18 · 2023: 23 · 2022: 29 · 2021: 33 · 2020: 21
- FWCI
- 6.87
- Type
- journal article
- Language
- en
- Update notices
- correction (2020-09-01)
- Open access
- gold
- doi.org
- Locations
- Nature Communications (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- National Institutes of Health
- National Heart, Lung, and Blood Institute
- NHLBI Division of Intramural Research
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Keywords
- histone modifications, single-cell epigenomics, Tn5 transposase, protein A, chromatin fragmentation, histone variants, chromatin-binding proteins
- MeSH terms
- Antibodies, Chromatin, Chromosome Mapping, Histones, Humans, Protein Processing, Post-Translational, Recombinant Fusion Proteins, Staining and Labeling, Staphylococcal Protein A, Sequence Analysis, DNA, Transposases, Histone Code, Chromatin Immunoprecipitation, HEK293 Cells, Epigenomics, Single-Cell Analysis
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