title: Targeted HIV-1 Latency Reversal Using CRISPR/Cas9-Derived Transcriptional Activator Systems.
abstract: CRISPR/Cas9 technology is currently considered the most advanced tool for targeted genome engineering. Its sequence-dependent specificity has been explored for locus-directed transcriptional modulation. Such modulation, in particular transcriptional activation, has been proposed as key approach to overcome silencing of dormant HIV provirus in latently infected cellular reservoirs. Currently available agents for provirus activation, so-called latency reversing agents (LRAs), act indirectly through cellular pathways to induce viral transcription. However, their clinical performance remains suboptimal, possibly because reservoirs have diverse cellular identities and/or proviral DNA is intractable to the induced pathways. We have explored two CRISPR/Cas9-derived activator systems as targeted approaches to induce dormant HIV-1 proviral DNA. These systems recruit multiple transcriptional activation domains to the HIV 5' long terminal repeat (LTR), for which we have identified an optimal target region within the LTR U3 sequence. Using this target region, we demonstrate transcriptional activation of proviral genomes via the synergistic activation mediator complex in various in culture model systems for HIV latency. Observed levels of induction are comparable or indeed higher than treatment with established LRAs. Importantly, activation is complete, leading to production of infective viral particles. Our data demonstrate that CRISPR/Cas9-derived technologies can be applied to counteract HIV latency and may therefore represent promising novel approaches in the quest for HIV elimination.
authors.affiliation: Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Julia K
authors.initials: JK
authors.lastname: Bialek
authors.affiliation: Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site, Hamburg, Germany.
authors.forename: Gábor A
authors.initials: GA
authors.lastname: Dunay
authors.affiliation: Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Maike
authors.initials: M
authors.lastname: Voges
authors.affiliation: Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site, Hamburg, Germany.
authors.forename: Carola
authors.initials: C
authors.lastname: Schäfer
authors.affiliation: Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Spohn
authors.affiliation: Friedrich-Baur-Institute, Department of Neurology, Ludwig Maximilian University Munich, Munich, Germany.
authors.forename: Rolf
authors.initials: R
authors.lastname: Stucka
authors.affiliation: Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site, Hamburg, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Hauber
authors.affiliation: Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.|Department of Anesthesiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.|German Center for Infection Research (DZIF), partner site, Hamburg, Germany.
authors.forename: Ulrike C
authors.initials: UC
authors.lastname: Lange
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D064113:CRISPR-Cas Systems; D000072669:Gene Editing; D018390:Gene Targeting; D015658:HIV Infections / Q000821:virology*; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D019169:Jurkat Cells; D011485:Protein Binding; D011533:Proviruses / Q000235:genetics; D000094704:RNA, Guide, CRISPR-Cas Systems; D015533:Transcriptional Activation; D017735:Virus Latency / Q000235:genetics; D014779:Virus Replication
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems