Live genome imaging by CRISPR engineering: progress and problems

Abstract CRISPR–Cas-based genome imaging opened a new era of genome visualization in living cells. While genomic loci with repetitive sequences, such as centromeres and telomeres, can be reliably imaged, applying the technique to nonrepetitive genomic loci has remained challenging. Recent advancemen...

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Main Authors: Eui-Jin Park, Hajin Kim
Format: Article
Language:English
Published: Nature Publishing Group 2025-07-01
Series:Experimental and Molecular Medicine
Online Access:https://doi.org/10.1038/s12276-025-01498-x
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author Eui-Jin Park
Hajin Kim
author_facet Eui-Jin Park
Hajin Kim
author_sort Eui-Jin Park
collection DOAJ
description Abstract CRISPR–Cas-based genome imaging opened a new era of genome visualization in living cells. While genomic loci with repetitive sequences, such as centromeres and telomeres, can be reliably imaged, applying the technique to nonrepetitive genomic loci has remained challenging. Recent advancements in the design of CRISPR RNAs and Cas proteins, the development of novel fluorophores and the combination of CRISPR–Cas with other molecular machinery amplified target-specific signals and suppressed background signals, revolutionizing this unique genome imaging technique and enabling the tracking of genomic loci with a small number of CRISPR–Cas complexes, down to a single complex. Here we review the latest advancements in CRISPR–Cas-based genome imaging techniques and their application to imaging nonrepetitive genomic loci. The challenges that these techniques are currently facing are the cellular toxicity and genomic instability induced by the expression of CRISPR–Cas and its interference with DNA metabolism, which impacts DNA replication and genome maintenance. Recently reported adverse effects of CRISPR–Cas-based genome labeling are discussed here, along with perspectives on how to overcome the problem.
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spelling doaj-art-e8342a42c0014855972d81f9ee2d57632025-08-20T04:01:43ZengNature Publishing GroupExperimental and Molecular Medicine2092-64132025-07-015771392139910.1038/s12276-025-01498-xLive genome imaging by CRISPR engineering: progress and problemsEui-Jin Park0Hajin Kim1Department of Biomedical Engineering, Ulsan National Institute of Science and TechnologyDepartment of Biomedical Engineering, Ulsan National Institute of Science and TechnologyAbstract CRISPR–Cas-based genome imaging opened a new era of genome visualization in living cells. While genomic loci with repetitive sequences, such as centromeres and telomeres, can be reliably imaged, applying the technique to nonrepetitive genomic loci has remained challenging. Recent advancements in the design of CRISPR RNAs and Cas proteins, the development of novel fluorophores and the combination of CRISPR–Cas with other molecular machinery amplified target-specific signals and suppressed background signals, revolutionizing this unique genome imaging technique and enabling the tracking of genomic loci with a small number of CRISPR–Cas complexes, down to a single complex. Here we review the latest advancements in CRISPR–Cas-based genome imaging techniques and their application to imaging nonrepetitive genomic loci. The challenges that these techniques are currently facing are the cellular toxicity and genomic instability induced by the expression of CRISPR–Cas and its interference with DNA metabolism, which impacts DNA replication and genome maintenance. Recently reported adverse effects of CRISPR–Cas-based genome labeling are discussed here, along with perspectives on how to overcome the problem.https://doi.org/10.1038/s12276-025-01498-x
spellingShingle Eui-Jin Park
Hajin Kim
Live genome imaging by CRISPR engineering: progress and problems
Experimental and Molecular Medicine
title Live genome imaging by CRISPR engineering: progress and problems
title_full Live genome imaging by CRISPR engineering: progress and problems
title_fullStr Live genome imaging by CRISPR engineering: progress and problems
title_full_unstemmed Live genome imaging by CRISPR engineering: progress and problems
title_short Live genome imaging by CRISPR engineering: progress and problems
title_sort live genome imaging by crispr engineering progress and problems
url https://doi.org/10.1038/s12276-025-01498-x
work_keys_str_mv AT euijinpark livegenomeimagingbycrisprengineeringprogressandproblems
AT hajinkim livegenomeimagingbycrisprengineeringprogressandproblems