A targeted CRISPR screen identifies ETS1 as a regulator of HIV-1 latency
Fig 9
ETS1 depletion increases the abundance of transcription activating histone marks at the HIV-1 LTR.
J-Lat 10.6 cells were nucleofected with ETS1 targeting or control non-targeting gRNAs. At 1 week post nucleofection, nucleofected cells were analyzed by Cleavage Under Targets & Release Using Nuclease (CUT&RUN) using antibodies against H3K9ac, H3K27ac and controls H3K4Me3 and IgG. (A) Experimental overview. (B) Western blot of the ETS1-targeting or control non-targeting (NT) gRNA nucleofected J-Lat10.6 cells for beta actin and ETS1 expression at one- week post nucleofection. (C) %GFP+ cells by flow cytometry 1 week post nucleofection with ETS1 and NT gRNA/Cas9 RNPs. (D) Abundance of Gag unspliced viral RNA expression in J-Lat 10.6 cells nucleofected with ETS1-targeting or NT control gRNA/Cas9 RNPs. Error bars represent standard deviation, and P values displayed were determined by two-way ANOVA Tukey’s multiple comparisons test. (E) Normalized coverage of sequencing reads (aggregated from 3 CUT&RUN technical replicates for each condition) across the HIV-1 reference genome and MALAT1 (chr11:65,498,907-65,506,539) is displayed using Integrative Genomics Viewer version 2.17. The HIV-1 LTR promoter region is denoted in a box. Figure elements prepared using Biorender.