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Isolated C. elegans germ nuclei exhibit distinct genomic profiles of histone modification and gene expression

BACKGROUND: The wide variety of specialized permissive and repressive mechanisms by which germ cells regulate developmental gene expression are not well understood genome-wide. Isolation of germ cells with high integrity and purity from living animals is necessary to address these open questions, bu...

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Autores principales: Han, Mei, Wei, Guifeng, McManus, Catherine E., Hillier, LaDeana W., Reinke, Valerie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6580472/
https://www.ncbi.nlm.nih.gov/pubmed/31208332
http://dx.doi.org/10.1186/s12864-019-5893-9
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author Han, Mei
Wei, Guifeng
McManus, Catherine E.
Hillier, LaDeana W.
Reinke, Valerie
author_facet Han, Mei
Wei, Guifeng
McManus, Catherine E.
Hillier, LaDeana W.
Reinke, Valerie
author_sort Han, Mei
collection PubMed
description BACKGROUND: The wide variety of specialized permissive and repressive mechanisms by which germ cells regulate developmental gene expression are not well understood genome-wide. Isolation of germ cells with high integrity and purity from living animals is necessary to address these open questions, but no straightforward methods are currently available. RESULTS: Here we present an experimental paradigm that permits the isolation of nuclei from C. elegans germ cells at quantities sufficient for genomic analyses. We demonstrate that these nuclei represent a very pure population and are suitable for both transcriptome analysis (RNA-seq) and chromatin immunoprecipitation (ChIP-seq) of histone modifications. From these data, we find unexpected germline- and soma-specific patterns of gene regulation. CONCLUSIONS: This new capacity removes a major barrier in the field to dissect gene expression mechanisms in the germ line of C. elegans. Consequent discoveries using this technology will be relevant to conserved regulatory mechanisms across species. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12864-019-5893-9) contains supplementary material, which is available to authorized users.
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spelling pubmed-65804722019-06-24 Isolated C. elegans germ nuclei exhibit distinct genomic profiles of histone modification and gene expression Han, Mei Wei, Guifeng McManus, Catherine E. Hillier, LaDeana W. Reinke, Valerie BMC Genomics Methodology Article BACKGROUND: The wide variety of specialized permissive and repressive mechanisms by which germ cells regulate developmental gene expression are not well understood genome-wide. Isolation of germ cells with high integrity and purity from living animals is necessary to address these open questions, but no straightforward methods are currently available. RESULTS: Here we present an experimental paradigm that permits the isolation of nuclei from C. elegans germ cells at quantities sufficient for genomic analyses. We demonstrate that these nuclei represent a very pure population and are suitable for both transcriptome analysis (RNA-seq) and chromatin immunoprecipitation (ChIP-seq) of histone modifications. From these data, we find unexpected germline- and soma-specific patterns of gene regulation. CONCLUSIONS: This new capacity removes a major barrier in the field to dissect gene expression mechanisms in the germ line of C. elegans. Consequent discoveries using this technology will be relevant to conserved regulatory mechanisms across species. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12864-019-5893-9) contains supplementary material, which is available to authorized users. BioMed Central 2019-06-17 /pmc/articles/PMC6580472/ /pubmed/31208332 http://dx.doi.org/10.1186/s12864-019-5893-9 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology Article
Han, Mei
Wei, Guifeng
McManus, Catherine E.
Hillier, LaDeana W.
Reinke, Valerie
Isolated C. elegans germ nuclei exhibit distinct genomic profiles of histone modification and gene expression
title Isolated C. elegans germ nuclei exhibit distinct genomic profiles of histone modification and gene expression
title_full Isolated C. elegans germ nuclei exhibit distinct genomic profiles of histone modification and gene expression
title_fullStr Isolated C. elegans germ nuclei exhibit distinct genomic profiles of histone modification and gene expression
title_full_unstemmed Isolated C. elegans germ nuclei exhibit distinct genomic profiles of histone modification and gene expression
title_short Isolated C. elegans germ nuclei exhibit distinct genomic profiles of histone modification and gene expression
title_sort isolated c. elegans germ nuclei exhibit distinct genomic profiles of histone modification and gene expression
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6580472/
https://www.ncbi.nlm.nih.gov/pubmed/31208332
http://dx.doi.org/10.1186/s12864-019-5893-9
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