Cited by 149Genomics and Chromatin DynamicsSingle-cell and spatial transcriptomicsEpigenetics and DNA Methylation

Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq)

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Carter, Benjamin, Ku, Wai Lim, Kang, Jee Youn, Hu, Gangqing, Perrie, Jonathan, Tang, Qingsong, Zhao, Keji·2019·Nature Communications

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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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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)
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
Open access
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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