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Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq)
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 fragm...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6702168/ https://www.ncbi.nlm.nih.gov/pubmed/31431618 http://dx.doi.org/10.1038/s41467-019-11559-1 |
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author | Carter, Benjamin Ku, Wai Lim Kang, Jee Youn Hu, Gangqing Perrie, Jonathan Tang, Qingsong Zhao, Keji |
author_facet | Carter, Benjamin Ku, Wai Lim Kang, Jee Youn Hu, Gangqing Perrie, Jonathan Tang, Qingsong Zhao, Keji |
author_sort | Carter, Benjamin |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-6702168 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-67021682019-08-22 Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq) Carter, Benjamin Ku, Wai Lim Kang, Jee Youn Hu, Gangqing Perrie, Jonathan Tang, Qingsong Zhao, Keji Nat Commun Article 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. Nature Publishing Group UK 2019-08-20 /pmc/articles/PMC6702168/ /pubmed/31431618 http://dx.doi.org/10.1038/s41467-019-11559-1 Text en © This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Carter, Benjamin Ku, Wai Lim Kang, Jee Youn Hu, Gangqing Perrie, Jonathan Tang, Qingsong Zhao, Keji Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq) |
title | Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq) |
title_full | Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq) |
title_fullStr | Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq) |
title_full_unstemmed | Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq) |
title_short | Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq) |
title_sort | mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (act-seq) |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6702168/ https://www.ncbi.nlm.nih.gov/pubmed/31431618 http://dx.doi.org/10.1038/s41467-019-11559-1 |
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