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In a screen for novel LRAs, we utilized fungal additional metabolites as a source of bioactive particles. Making use of orthogonal mass spectrometry (MS) paired to latency reversal bioassays, we identified gliotoxin (GTX) as a novel LRA. GTX significantly caused HIV-1 gene phrase in latent ex vivo infected primary cells and in CD4+ T cells from all aviremic HIV-1+ individuals. RNA sequencing identified 7SK RNA, the scaffold of the positive transcription elongation