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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...

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Autores principales: Carter, Benjamin, Ku, Wai Lim, Kang, Jee Youn, Hu, Gangqing, Perrie, Jonathan, Tang, Qingsong, Zhao, Keji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
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.
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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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