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A streamlined tethered chromosome conformation capture protocol

BACKGROUND: Identification of locus-locus contacts at the chromatin level provides a valuable foundation for understanding of nuclear architecture and function and a valuable tool for inferring long-range linkage relationships. As one approach to this, chromatin conformation capture-based techniques...

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Autores principales: Gabdank, Idan, Ramakrishnan, Sreejith, Villeneuve, Anne M., Fire, Andrew Z.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4818521/
https://www.ncbi.nlm.nih.gov/pubmed/27036078
http://dx.doi.org/10.1186/s12864-016-2596-3
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author Gabdank, Idan
Ramakrishnan, Sreejith
Villeneuve, Anne M.
Fire, Andrew Z.
author_facet Gabdank, Idan
Ramakrishnan, Sreejith
Villeneuve, Anne M.
Fire, Andrew Z.
author_sort Gabdank, Idan
collection PubMed
description BACKGROUND: Identification of locus-locus contacts at the chromatin level provides a valuable foundation for understanding of nuclear architecture and function and a valuable tool for inferring long-range linkage relationships. As one approach to this, chromatin conformation capture-based techniques allow creation of genome spatial organization maps. While such approaches have been available for some time, methodological advances will be of considerable use in minimizing both time and input material required for successful application. RESULTS: Here we report a modified tethered conformation capture protocol that utilizes a series of rapid and efficient molecular manipulations. We applied the method to Caenorhabditis elegans, obtaining chromatin interaction maps that provide a sequence-anchored delineation of salient aspects of Caenorhabditis elegans chromosome structure, demonstrating a high level of consistency in overall chromosome organization between biological samples collected under different conditions. In addition to the application of the method to defining nuclear architecture, we found the resulting chromatin interaction maps to be of sufficient resolution and sensitivity to enable detection of large-scale structural variants such as inversions or translocations. CONCLUSION: Our streamlined protocol provides an accelerated, robust, and broadly applicable means of generating chromatin spatial organization maps and detecting genome rearrangements without a need for cellular or chromatin fractionation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-016-2596-3) contains supplementary material, which is available to authorized users.
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spelling pubmed-48185212016-04-03 A streamlined tethered chromosome conformation capture protocol Gabdank, Idan Ramakrishnan, Sreejith Villeneuve, Anne M. Fire, Andrew Z. BMC Genomics Methodology Article BACKGROUND: Identification of locus-locus contacts at the chromatin level provides a valuable foundation for understanding of nuclear architecture and function and a valuable tool for inferring long-range linkage relationships. As one approach to this, chromatin conformation capture-based techniques allow creation of genome spatial organization maps. While such approaches have been available for some time, methodological advances will be of considerable use in minimizing both time and input material required for successful application. RESULTS: Here we report a modified tethered conformation capture protocol that utilizes a series of rapid and efficient molecular manipulations. We applied the method to Caenorhabditis elegans, obtaining chromatin interaction maps that provide a sequence-anchored delineation of salient aspects of Caenorhabditis elegans chromosome structure, demonstrating a high level of consistency in overall chromosome organization between biological samples collected under different conditions. In addition to the application of the method to defining nuclear architecture, we found the resulting chromatin interaction maps to be of sufficient resolution and sensitivity to enable detection of large-scale structural variants such as inversions or translocations. CONCLUSION: Our streamlined protocol provides an accelerated, robust, and broadly applicable means of generating chromatin spatial organization maps and detecting genome rearrangements without a need for cellular or chromatin fractionation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-016-2596-3) contains supplementary material, which is available to authorized users. BioMed Central 2016-04-01 /pmc/articles/PMC4818521/ /pubmed/27036078 http://dx.doi.org/10.1186/s12864-016-2596-3 Text en © Gabdank et al. 2016 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
Gabdank, Idan
Ramakrishnan, Sreejith
Villeneuve, Anne M.
Fire, Andrew Z.
A streamlined tethered chromosome conformation capture protocol
title A streamlined tethered chromosome conformation capture protocol
title_full A streamlined tethered chromosome conformation capture protocol
title_fullStr A streamlined tethered chromosome conformation capture protocol
title_full_unstemmed A streamlined tethered chromosome conformation capture protocol
title_short A streamlined tethered chromosome conformation capture protocol
title_sort streamlined tethered chromosome conformation capture protocol
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4818521/
https://www.ncbi.nlm.nih.gov/pubmed/27036078
http://dx.doi.org/10.1186/s12864-016-2596-3
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