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Micro-C XL: assaying chromosome conformation at length scales from the nucleosome to the entire genome

Tsung-Han S. Hsieh, Geoffrey Fudenberg, Anton Goloborodko, Oliver J. Rando
doi: https://doi.org/10.1101/071357
Tsung-Han S. Hsieh
1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA
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Geoffrey Fudenberg
2Department of Physics and Center for 3D Structure and Physics of the Genome, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA
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Anton Goloborodko
2Department of Physics and Center for 3D Structure and Physics of the Genome, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA
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Oliver J. Rando
1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA
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  • For correspondence: Oliver.Rando@umassmed.edu
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ABSTRACT

Structural analysis of chromosome folding in vivo has been revolutionized by Chromosome Conformation Capture (3C) and related methods, which use proximity ligation to identify chromosomal loci in physical contact. We recently described a variant 3C technique, Micro-C, in which chromatin is fragmented to mononucleosomes using micrococcal nuclease, enabling nucleosome-resolution folding maps of the genome. Here, we describe an improved Micro-C protocol using long crosslinkers, termed Micro-C XL, which exhibits greatly increased signal to noise, and provides further insight into the folding of the yeast genome. We also find that signal to noise is much improved in Micro-C XL libraries generated from relatively insoluble chromatin as opposed to soluble material, providing a simple method to physically enrich for bona-fide long-range interactions. Micro-C XL maps of the budding and fission yeast genomes reveal both short-range chromosome fiber features such as chromosomally-interacting domains (CIDs), as well as higher-order features such as clustering of centromeres and telomeres, thereby addressing the primary discrepancy between prior Micro-C data and reported 3C and Hi-C analyses. Interestingly, comparison of chromosome folding maps of S. cerevisiae and S. pombe revealed widespread qualitative similarities, yet quantitative differences, between these distantly-related species. Micro-C XL thus provides a single assay suitable for interrogation of chromosome folding at length scales from the nucleosome to the full genome.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted August 24, 2016.
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Micro-C XL: assaying chromosome conformation at length scales from the nucleosome to the entire genome
Tsung-Han S. Hsieh, Geoffrey Fudenberg, Anton Goloborodko, Oliver J. Rando
bioRxiv 071357; doi: https://doi.org/10.1101/071357
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Micro-C XL: assaying chromosome conformation at length scales from the nucleosome to the entire genome
Tsung-Han S. Hsieh, Geoffrey Fudenberg, Anton Goloborodko, Oliver J. Rando
bioRxiv 071357; doi: https://doi.org/10.1101/071357

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