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CRISPR-Based DNA Imaging in Living Cells Reveals Cell Cycle-Dependent Chromosome Dynamics

Hanhui Ma, Li-Chun Tu, Ardalan Naseri, Yu-Chieh Chung, David Grunwald, Shaojie Zhang, Thoru Pederson
doi: https://doi.org/10.1101/195966
Hanhui Ma
1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA
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  • For correspondence: hanhui.ma@umassmed.edu
Li-Chun Tu
2RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USA
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Ardalan Naseri
3Department of Computer Science, University of Central Florida, Orlando, FL 32816, USA
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Yu-Chieh Chung
4Kavli Institute for the Physics and Mathematics of the Universe, University of Tokyo, Kashiwa, Chiba, 277-8583, Japan
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David Grunwald
2RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USA
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Shaojie Zhang
3Department of Computer Science, University of Central Florida, Orlando, FL 32816, USA
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Thoru Pederson
1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA
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SUMMARY

In contrast to the well-studied condensation and folding of chromosomes during mitosis, their dynamics in interphase are less understood. We developed a sensitive, multicolor system, CRISPR-Sirius, allowing the real-time tracking of the dynamics of chromosomal loci. We tracked loci kilobases to megabases apart and found significant variation in the inter-locus distances of each pair, indicating differing degrees of DNA contortion. We resolved two distinct modes of dynamics of loci: saltatory local movements as well as translational movements of the domain. The magnitude of both of these modes of movements increased from early to late G1, whereas the translational movements were reduced in early S. The local fluctuations decreased slightly in early S and more markedly in mid-late S. These newly observed movements and their cell cycle-dependence are indicative of a hitherto unrecognized compaction-relaxation dynamic of the chromosomal fiber operating concurrently with changes in the extent of observed genomic domain movements.

IN BRIEF Distinct chromosome folding and dynamics during cell cycle progression were dissected by CRISPR-Sirius DNA imaging in living cells.

HIGHLIGHTS

  • CRISPR-Sirius allows tracking of pairs of chromosomal loci having kilobase to megabase inter-locus distances

  • Pair-wise tracking of loci allows measurement of both local and domain dynamics

  • Chromosomal fiber relaxation is positively correlated with local dynamics

  • Genomic region size contributes to local and domain movements

  • Distinct chromosome dynamics were uncovered during cell cycle progression in interphase

Copyright 
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-ND 4.0 International license.
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Posted September 29, 2017.
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CRISPR-Based DNA Imaging in Living Cells Reveals Cell Cycle-Dependent Chromosome Dynamics
Hanhui Ma, Li-Chun Tu, Ardalan Naseri, Yu-Chieh Chung, David Grunwald, Shaojie Zhang, Thoru Pederson
bioRxiv 195966; doi: https://doi.org/10.1101/195966
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CRISPR-Based DNA Imaging in Living Cells Reveals Cell Cycle-Dependent Chromosome Dynamics
Hanhui Ma, Li-Chun Tu, Ardalan Naseri, Yu-Chieh Chung, David Grunwald, Shaojie Zhang, Thoru Pederson
bioRxiv 195966; doi: https://doi.org/10.1101/195966

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