A targeted CRISPR screen identifies ETS1 as a regulator of HIV-1 latency
Fig 6
ETS1 knockout reactivates HIV-1 transcription in CD4 T cells from people with HIV-1 (PWH) on antiretroviral therapy (ART) ex vivo.
(A) Schematic of ex vivo CRISPR nucleofection experiment in resting CD4 T cells isolated from PWH on ART. CRISPR nucleofection of ETS1-targeting or non-targeting (NT) Cas9/gRNAs was performed after a 24 h culture of PWH CD4 T cells followed by quantification of Gag RNA expression at 3 days post nucleofection by quantitative PCR. (B) Depletion of ETS1 in nucleofected resting CD4 T cells was analyzed by western blot. (C) Relative abundance of Gag viral RNA expression in CD4 T cells nucleofected with ETS1-targeting or NT control Cas9/gRNA complexes. Gag viral RNA expression was normalized to cells nucleofected with non-targeting gRNA and presented as fold change. Experiment was conducted in three biological replicates/donors with three technical replicates for each condition. Error bars represent standard deviation, and P values displayed were determined by two-way ANOVA Tukey’s multiple comparisons test. Figure elements prepared using Biorender.