Disruption of the Integrator complex triggers the Integrated Stress Response through formation of double-stranded RNA [PROseq]

Acute loss of INT induces transcription factors that initiate stress-responses like the Integrated Stress Response (ISR). Inefficient elongation upon INT loss enables termination in introns and dsRNA formation. dsRNA is recognized by PKR, which persistently activates the ISR in samples from patients with INT mutations PROseq performed in presence or absence of acute degradation of INTS1 in HEK293T cells. Cells were permeabilized as in Mimoso et al. 2024; run-ons were performed with biotinylated NTPs and then purified. Purified biotinylated RNAs were ligated to sequencing adapters and amplified by PCR.