A targeted CRISPR screen identifies ETS1 as a regulator of HIV-1 latency
Fig 8
Reactivation of HIV-1 following ETS1 depletion is partially dependent on MALAT1.
In vitro generated primary latent CD4 T cells were nucleofected with Cas9/gRNA ribonucleoprotein complexes targeting ETS1, MALAT1, or a combination of ETS1 and MALAT1 as described in Fig 6, followed by immunoblotting, flow cytometry and RT-PCR. (A) Depletion of ETS1 in nucleofected CD4 T cells was analyzed by western blot. (B-D) Relative abundance of MALAT1 RNA expression (B), and Gag RNA (C), and HIV-1 protein expression (D) in CD4 T cells nucleofected with Cas9/sgRNAs targeting ETS1, MALAT1, or a combination of ETS1 and MALAT1. MALAT1 and Gag viral RNA expression was normalized to cells nucleofected with NT control. Experiment was conducted in three biological replicates/donors with three technical replicates for each condition. Error bars represent standard deviations, and P values displayed were determined by two-way ANOVA Tukey’s multiple comparisons test.