title: Elimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing.
abstract: 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.
authors.affiliation: Department of Neuroscience/Center for Neurovirology, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.|Comprehensive NeuroAIDS Center, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.
authors.forename: Rafal
authors.initials: R
authors.lastname: Kaminski
authors.affiliation: Department of Neuroscience/Center for Neurovirology, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.|Comprehensive NeuroAIDS Center, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.
authors.forename: Yilan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Neuroscience/Center for Neurovirology, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.|Comprehensive NeuroAIDS Center, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.
authors.forename: Tracy
authors.initials: T
authors.lastname: Fischer
authors.affiliation: Comprehensive NeuroAIDS Center, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.|Department of Medicine, Temple HIV Program, Lewis Katz School of Medicine at Temple University/Temple University Hospital, 3400 N. Broad Street, Philadelphia, PA 19140 USA.
authors.forename: Ellen
authors.initials: E
authors.lastname: Tedaldi
authors.affiliation: Department of Neuroscience/Center for Neurovirology, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.|Comprehensive NeuroAIDS Center, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Napoli
authors.affiliation: Department of Neuroscience/Center for Neurovirology, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.|Comprehensive NeuroAIDS Center, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, OH 44106, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Karn
authors.affiliation: Department of Neuroscience/Center for Neurovirology, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.|Comprehensive NeuroAIDS Center, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.
authors.forename: Wenhui
authors.initials: W
authors.lastname: Hu
authors.affiliation: Department of Neuroscience/Center for Neurovirology, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.|Comprehensive NeuroAIDS Center, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140 USA.
authors.forename: Kamel
authors.initials: K
authors.lastname: Khalili
mesh_terms: D017209:Apoptosis; D001426:Bacterial Proteins / Q000235:genetics*; D015496:CD4-Positive T-Lymphocytes / Q000502:physiology / Q000821:virology*; D000076987:CRISPR-Associated Protein 9; D064113:CRISPR-Cas Systems / Q000235:genetics*; D002453:Cell Cycle; D002470:Cell Survival; D004720:Endonucleases / Q000235:genetics*; D000072669:Gene Editing; D015316:Genetic Therapy; D015658:HIV Infections / Q000628:therapy*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D019169:Jurkat Cells; D011533:Proviruses / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D017735:Virus Latency / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D012367:RNA, Viral; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases