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유전체 항상성 연구단
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Background-suppressed live visualization of genomic loci with an improved CRISPR system based on a split fluorophore

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Title
Background-suppressed live visualization of genomic loci with an improved CRISPR system based on a split fluorophore
Author(s)
Narendra Chaudhary; Si-Hyeong Nho; Hayoon Cho; Narangerel Gantumur; Jae Sun Ra; Kyungjae Myung; Hajin Kim
Subject
REPETITIVE DNA-SEQUENCES, ; TRANSLATION DYNAMICS, ; HUMAN-CELLS, ; ORGANIZATION, ; PROTEIN, ; LOCALIZATION, ; BINDING, ; DESIGN, ; SGRNAS, ; CAS9
Publication Date
2020-09
Journal
GENOME RESEARCH, v.30, no.9, pp.1306 - 1316
Publisher
Cold Spring Harbor Laboratory Press
Abstract
© 2020 Chaudhary et al. The higher-order structural organization and dynamics of the chromosomes play a central role in gene regulation. To explore this structure-function relationship, it is necessary to directly visualize genomic elements in living cells. Genome imaging based on the CRISPR system is a powerful approach but has limited applicability due to background signals and nonspecific aggregation of fluorophores within nuclei. To address this issue, we developed a novel visualization scheme combining tripartite fluorescent proteins with the SunTag system and demonstrated that it strongly suppressed background fluorescence and amplified locus-specific signals, allowing long-term tracking of genomic loci. We integrated the multicomponent CRISPR system into stable cell lines to allow quantitative and reliable analysis of dynamic behaviors of genomic loci. Due to the greatly elevated signal-to-background ratio, target loci with only small numbers of sequence repeats could be successfully tracked, even under a conventional fluorescence microscope. This feature enables the application of CRISPR-based imaging to loci throughout the genome and opens up new possibilities for the study of nuclear processes in living cells
URI
https://pr.ibs.re.kr/handle/8788114/7630
DOI
10.1101/GR.260018.119
ISSN
1088-9051
Appears in Collections:
Center for Genomic Integrity(유전체 항상성 연구단) > 1. Journal Papers (저널논문)
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