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Kim, Hajin
Single Molecule Biophysics Lab
Research Interests
  • Biophysics, Single Molecule, fluorescence Spectroscopy, biological Motors, biomolecular assembly

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Visualizing Live Chromatin Dynamics through CRISPR-Based Imaging Techniques

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Title
Visualizing Live Chromatin Dynamics through CRISPR-Based Imaging Techniques
Author
Chaudhary, NarendraIm, Jae-KyeongNho, Si-HyeongKim, Hajin
Issue Date
2021-09
Publisher
KOREAN SOC MOLECULAR & CELLULAR BIOLOGY
Citation
MOLECULES AND CELLS, v.44, no.9, pp.627 - 636
Abstract
The three-dimensional organization of chromatin and its time-dependent changes greatly affect virtually every cellular function, especially DNA replication, genome maintenance, transcription regulation, and cell differentiation. Sequencing-based techniques such as ChIP-seq, ATAC-seq, and Hi-C provide abundant information on how genomic elements are coupled with regulatory proteins and functionally organized into hierarchical domains through their interactions. However, visualizing the time-dependent changes of such organization in individual cells remains challenging. Recent developments of CRISPR systems for site-specific fluorescent labeling of genomic loci have provided promising strategies for visualizing chromatin dynamics in live cells. However, there are several limiting factors, including background signals, off-target binding of CRISPR, and rapid photobleaching of the fluorophores, requiring a large number of target-bound CRISPR complexes to reliably distinguish the target-specific foci from the background. Various modifications have been engineered into the CRISPR system to enhance the signal-to-background ratio and signal longevity to detect target foci more reliably and efficiently, and to reduce the required target size. In this review, we comprehensively compare the performances of recently developed CRISPR designs for improved visualization of genomic loci in terms of the reliability of target detection, the ability to detect small repeat loci, and the allowed time of live tracking. Longer observation of genomic loci allows the detailed identification of the dynamic characteristics of chromatin. The diffusion properties of chromatin found in recent studies are reviewed, which provide suggestions for the underlying biological processes.
URI
https://scholarworks.unist.ac.kr/handle/201301/54832
URL
https://www.molcells.org/journal/view.html?doi=10.14348/molcells.2021.2254
DOI
10.14348/molcells.2021.2254
ISSN
1016-8478
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BME_Journal Papers
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