A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1
The relative positions of viral DNA genomes to the host intranuclear environment play critical roles in determining virus fate. Recent advances in the application of chromosome conformation capture-based sequencing analysis (3 C technologies) have revealed valuable aspects of the spatiotemporal inte...
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eLife Sciences Publications Ltd
2023-09-01
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| Online Access: | https://elifesciences.org/articles/85412 |
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| author | Juan Xiang Chaoyang Fan Hongchang Dong Yilei Ma Pei Xu |
| author_facet | Juan Xiang Chaoyang Fan Hongchang Dong Yilei Ma Pei Xu |
| author_sort | Juan Xiang |
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| description | The relative positions of viral DNA genomes to the host intranuclear environment play critical roles in determining virus fate. Recent advances in the application of chromosome conformation capture-based sequencing analysis (3 C technologies) have revealed valuable aspects of the spatiotemporal interplay of viral genomes with host chromosomes. However, to elucidate the causal relationship between the subnuclear localization of viral genomes and the pathogenic outcome of an infection, manipulative tools are needed. Rapid repositioning of viral DNAs to specific subnuclear compartments amid infection is a powerful approach to synchronize and interrogate this dynamically changing process in space and time. Herein, we report an inducible CRISPR-based two-component platform that relocates extrachromosomal DNA pieces (5 kb to 170 kb) to the nuclear periphery in minutes (CRISPR-nuPin). Based on this strategy, investigations of herpes simplex virus 1 (HSV-1), a prototypical member of the human herpesvirus family, revealed unprecedently reported insights into the early intranuclear life of the pathogen: (I) Viral genomes tethered to the nuclear periphery upon entry, compared with those freely infecting the nucleus, were wrapped around histones with increased suppressive modifications and subjected to stronger transcriptional silencing and prominent growth inhibition. (II) Relocating HSV-1 genomes at 1 hr post infection significantly promoted the transcription of viral genes, termed an ‘Escaping’ effect. (III) Early accumulation of ICP0 was a sufficient but not necessary condition for ‘Escaping’. (IV) Subnuclear localization was only critical during early infection. Importantly, the CRISPR-nuPin tactic, in principle, is applicable to many other DNA viruses. |
| format | Article |
| id | doaj-art-24c29dba6e6c44849ad2c220abf333e9 |
| institution | Kabale University |
| issn | 2050-084X |
| language | English |
| publishDate | 2023-09-01 |
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| spelling | doaj-art-24c29dba6e6c44849ad2c220abf333e92024-11-11T16:57:12ZengeLife Sciences Publications LtdeLife2050-084X2023-09-011210.7554/eLife.85412A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1Juan Xiang0https://orcid.org/0000-0003-3853-7762Chaoyang Fan1https://orcid.org/0000-0003-2286-4865Hongchang Dong2https://orcid.org/0000-0002-8190-2158Yilei Ma3Pei Xu4https://orcid.org/0000-0003-2719-5605The Centre for Infection and Immunity Studies, School of Medicine, Shenzhen Campus of Sun Yat-sen Univeristy, Sun Yat-sen University, Shenzhen, ChinaThe Centre for Infection and Immunity Studies, School of Medicine, Shenzhen Campus of Sun Yat-sen Univeristy, Sun Yat-sen University, Shenzhen, ChinaThe Centre for Infection and Immunity Studies, School of Medicine, Shenzhen Campus of Sun Yat-sen Univeristy, Sun Yat-sen University, Shenzhen, ChinaThe Centre for Infection and Immunity Studies, School of Medicine, Shenzhen Campus of Sun Yat-sen Univeristy, Sun Yat-sen University, Shenzhen, ChinaThe Centre for Infection and Immunity Studies, School of Medicine, Shenzhen Campus of Sun Yat-sen Univeristy, Sun Yat-sen University, Shenzhen, ChinaThe relative positions of viral DNA genomes to the host intranuclear environment play critical roles in determining virus fate. Recent advances in the application of chromosome conformation capture-based sequencing analysis (3 C technologies) have revealed valuable aspects of the spatiotemporal interplay of viral genomes with host chromosomes. However, to elucidate the causal relationship between the subnuclear localization of viral genomes and the pathogenic outcome of an infection, manipulative tools are needed. Rapid repositioning of viral DNAs to specific subnuclear compartments amid infection is a powerful approach to synchronize and interrogate this dynamically changing process in space and time. Herein, we report an inducible CRISPR-based two-component platform that relocates extrachromosomal DNA pieces (5 kb to 170 kb) to the nuclear periphery in minutes (CRISPR-nuPin). Based on this strategy, investigations of herpes simplex virus 1 (HSV-1), a prototypical member of the human herpesvirus family, revealed unprecedently reported insights into the early intranuclear life of the pathogen: (I) Viral genomes tethered to the nuclear periphery upon entry, compared with those freely infecting the nucleus, were wrapped around histones with increased suppressive modifications and subjected to stronger transcriptional silencing and prominent growth inhibition. (II) Relocating HSV-1 genomes at 1 hr post infection significantly promoted the transcription of viral genes, termed an ‘Escaping’ effect. (III) Early accumulation of ICP0 was a sufficient but not necessary condition for ‘Escaping’. (IV) Subnuclear localization was only critical during early infection. Importantly, the CRISPR-nuPin tactic, in principle, is applicable to many other DNA viruses.https://elifesciences.org/articles/85412intranuclear positioningHSV-1heterochromatin packagingCRISPR/dcas9DNA virus |
| spellingShingle | Juan Xiang Chaoyang Fan Hongchang Dong Yilei Ma Pei Xu A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1 eLife intranuclear positioning HSV-1 heterochromatin packaging CRISPR/dcas9 DNA virus |
| title | A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1 |
| title_full | A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1 |
| title_fullStr | A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1 |
| title_full_unstemmed | A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1 |
| title_short | A CRISPR-based rapid DNA repositioning strategy and the early intranuclear life of HSV-1 |
| title_sort | crispr based rapid dna repositioning strategy and the early intranuclear life of hsv 1 |
| topic | intranuclear positioning HSV-1 heterochromatin packaging CRISPR/dcas9 DNA virus |
| url | https://elifesciences.org/articles/85412 |
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