Genome-wide CRISPR/Cas9 screening to identify novel response regulators to double-stranded RNA stress in lung adenocarcinoma

Abstract

Double-stranded RNAs (dsRNAs) are key players in the innate immune response to viral infection. Upon infection, dsRNAs generated during viral transcription or the dsRNA viral genome itself are recognized by pattern recognition receptors (PRRs) on host cells, such as Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), and cytosolic dsRNA sensors. This recognition leads to the activation of the downstream antiviral signaling pathways, including the production of type I interferons (IFNs) and pro-inflammatory cytokines and the induction of antiviral genes. However, the mitochondrial genome and the transposable elements also produce long dsRNAs that are closely associated with disease pathogenesis by triggering an aberrant immune response. The importance of dsRNA-mediated immune activation has led to the identification of various downstream regulators of the dsRNA stress response, such as PAMP receptors and dsRNA-binding proteins. However, an unbiased systematic approach to determine the key regulators and elucidate their regulatory mechanisms for dsRNAs is currently lacking. Hence, we perform a genome-wide CRISPR/Cas9 knockout screening using poly I:C as a dsRNA-mimicking stressor. Our screening hits validated previously known dsRNA receptors and regulators of inflammatory response. Interestingly, we also identified protein such as LSM12 that may play essential roles in regulating the dsRNA-mediated stress response. Our study aims to elucidate these regulators and their regulatory mechanism of innate immune response to dsRNAs.