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Zhang Y , Yin C , Zhang T , Li F , Yang W , Kaminski R , Fagan PR , Putatunda R , Young WB , Khalili K , Hu W
CRISPR/gRNA-directed synergistic activation mediator (SAM) induces specific, persistent and robust reactivation of the HIV-1 latent reservoirs
Sci Rep. 2015 ;5 :16277
PMID: 26538064    PMCID: PMC4633726   
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Current antiretroviral therapy does not eliminate the integrated and transcriptionally silent HIV-1 provirus in latently infected cells. Recently, a "shock and kill" strategy has been extensively explored to eradicate the HIV-1 latent reservoirs for a permanent cure of AIDS. The therapeutic efficacy of currently used agents remains disappointing because of low efficiency, non-specificity and cellular toxicity. Here we present a novel catalytically-deficient Cas9-synergistic activation mediator (dCas9-SAM) technology to selectively, potently and persistently reactivate the HIV-1 latent reservoirs. By screening 16 MS2-mediated single guide RNAs, we identified long terminal repeat (LTR)-L and O that surround the enhancer region (-165/-145 for L and -92/-112 for O) and induce robust reactivation of HIV-1 provirus in HIV-1 latent TZM-bI epithelial, Jurkat T lymphocytic and CHME5 microglial cells. This compulsory reactivation induced cellular suicide via toxic buildup of viral proteins within HIV-1 latent Jurkat T and CHME5 microglial cells. These results suggest that this highly effective and target-specific dCas9-SAM system can serve as a novel HIV-latency-reversing therapeutic tool for the permanent elimination of HIV-1 latent reservoirs.
Zhang, Yonggang Yin, Chaoran Zhang, Ting Li, Fang Yang, Wensheng Kaminski, Rafal Fagan, Philip Regis Putatunda, Raj Young, Won-Bin Khalili, Kamel Hu, Wenhui P30 MH092177/MH/NIMH NIH HHS/United States R01 DK075964/DK/NIDDK NIH HHS/United States R01 MH101041/MH/NIMH NIH HHS/United States R01 NS087971/NS/NINDS NIH HHS/United States England Sci Rep. 2015 Nov 5;5:16277. doi: 10.1038/srep16277.