A targeted CRISPR screen identifies ETS1 as a regulator of HIV-1 latency
Fig 4
ETS1 knockout in primary CD4 T cells reveals positive and negative roles in HIV-1 expression.
Primary CD4 T cells were activated, infected with HIV-GFP, enriched and nucleofected as in Fig 3. (A) Western blot of the ETS1-targeting or a control non-targeting (NT) Cas9/gRNA nucleofected primary cells for beta actin and ETS1 expression at one and two weeks post nucleofection (pKO). (B-E) Flow cytometry of infected cells without stimulation or after 24 h of stimulation with one of four different LRAs (vorinostat, prostratin, iBET151, or AZD5582) at one (B) and two (D) week post nucleofection. (C and E) Dot plots representing viral GFP expression at one and two weeks post knockout respectively. Data are represented as fold change in percent GFP expression, normalized to cells nucleofected with NT gRNA. Error bars represent standard deviation, and P values displayed were determined by two-way ANOVA Tukey’s multiple comparisons test. Data shown are representative of two independent experiments with different donors, with biological triplicates for each condition.