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Elimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing.

We employed an RNA-guided CRISPR/Cas9 DNA editing system to precisely remove the entire HIV-1 genome spanning between 5' and 3' LTRs of integrated HIV-1 proviral DNA copies from latently infected human CD4+ T-cells. Comprehensive assessment of whole-genome sequencing of HIV-1 eradicated cells ruled... Full description

Journal Title: Scientific reports March 4, 2016, Vol.6, p.22555
Main Author: Kaminski, Rafal
Other Authors: Chen, Yilan , Fischer, Tracy , Tedaldi, Ellen , Napoli, Alessandro , Zhang, Yonggang , Karn, Jonathan , Hu, Wenhui , Khalili, Kamel
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: E-ISSN: 2045-2322 ; DOI: 10.1038/srep22555
Link: http://search.proquest.com/docview/1770873115/?pq-origsite=primo
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recordid: proquest1770873115
title: Elimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing.
format: Article
creator:
  • Kaminski, Rafal
  • Chen, Yilan
  • Fischer, Tracy
  • Tedaldi, Ellen
  • Napoli, Alessandro
  • Zhang, Yonggang
  • Karn, Jonathan
  • Hu, Wenhui
  • Khalili, Kamel
subjects:
  • Apoptosis–Genetics
  • Bacterial Proteins–Physiology
  • Cd4-Positive T-Lymphocytes–Virology
  • Crispr-Associated Protein 9–Genetics
  • Crispr-Cas Systems–Genetics
  • Cell Cycle–Therapy
  • Cell Survival–Genetics
  • Endonucleases–Genetics
  • Gene Editing–Genetics
  • Genetic Therapy–Genetics
  • HIV Infections–Genetics
  • HIV-1–Genetics
  • Humans–Genetics
  • Jurkat Cells–Genetics
  • Proviruses–Genetics
  • RNA, Viral–Genetics
  • Virus Latency–Genetics
  • Bacterial Proteins
  • RNA, Viral
  • Crispr-Associated Protein 9
ispartof: Scientific reports, March 4, 2016, Vol.6, p.22555
description: We employed an RNA-guided CRISPR/Cas9 DNA editing system to precisely remove the entire HIV-1 genome spanning between 5' and 3' LTRs of integrated HIV-1 proviral DNA copies from latently infected human CD4+ T-cells. Comprehensive assessment of whole-genome sequencing of HIV-1 eradicated cells ruled out any off-target effects by our CRISPR/Cas9 technology that might compromise the integrity of the host genome and further showed no effect on several cell health indices including viability, cell cycle and apoptosis. Persistent co-expression of Cas9 and the specific targeting guide RNAs in HIV-1-eradicated T-cells protected them against new infection by HIV-1. Lentivirus-delivered CRISPR/Cas9 significantly diminished HIV-1 replication in infected primary CD4+ T-cell cultures and drastically reduced viral load in ex vivo culture of CD4+ T-cells obtained from HIV-1 infected patients. Thus, gene editing using CRISPR/Cas9 may provide a new therapeutic path for eliminating HIV-1 DNA from CD4+ T-cells and potentially serve as a novel and effective platform toward curing AIDS.
language: eng
source:
identifier: E-ISSN: 2045-2322 ; DOI: 10.1038/srep22555
fulltext: fulltext
issn:
  • 20452322
  • 2045-2322
url: Link


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titleElimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing.
creatorKaminski, Rafal ; Chen, Yilan ; Fischer, Tracy ; Tedaldi, Ellen ; Napoli, Alessandro ; Zhang, Yonggang ; Karn, Jonathan ; Hu, Wenhui ; Khalili, Kamel
contributorKaminski, Rafal (correspondence author) ; Kaminski, Rafal (record owner)
ispartofScientific reports, March 4, 2016, Vol.6, p.22555
identifierE-ISSN: 2045-2322 ; DOI: 10.1038/srep22555
subjectApoptosis–Genetics ; Bacterial Proteins–Physiology ; Cd4-Positive T-Lymphocytes–Virology ; Crispr-Associated Protein 9–Genetics ; Crispr-Cas Systems–Genetics ; Cell Cycle–Therapy ; Cell Survival–Genetics ; Endonucleases–Genetics ; Gene Editing–Genetics ; Genetic Therapy–Genetics ; HIV Infections–Genetics ; HIV-1–Genetics ; Humans–Genetics ; Jurkat Cells–Genetics ; Proviruses–Genetics ; RNA, Viral–Genetics ; Virus Latency–Genetics ; Bacterial Proteins ; RNA, Viral ; Crispr-Associated Protein 9
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descriptionWe employed an RNA-guided CRISPR/Cas9 DNA editing system to precisely remove the entire HIV-1 genome spanning between 5' and 3' LTRs of integrated HIV-1 proviral DNA copies from latently infected human CD4+ T-cells. Comprehensive assessment of whole-genome sequencing of HIV-1 eradicated cells ruled out any off-target effects by our CRISPR/Cas9 technology that might compromise the integrity of the host genome and further showed no effect on several cell health indices including viability, cell cycle and apoptosis. Persistent co-expression of Cas9 and the specific targeting guide RNAs in HIV-1-eradicated T-cells protected them against new infection by HIV-1. Lentivirus-delivered CRISPR/Cas9 significantly diminished HIV-1 replication in infected primary CD4+ T-cell cultures and drastically reduced viral load in ex vivo culture of CD4+ T-cells obtained from HIV-1 infected patients. Thus, gene editing using CRISPR/Cas9 may provide a new therapeutic path for eliminating HIV-1 DNA from CD4+ T-cells and potentially serve as a novel and effective platform toward curing AIDS.
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titleElimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing.
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titleElimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing.
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