IRGQ Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the IRGQ gene in the HAP1 host cell line. This near-haploid human fibroblast-like cell model enables loss-of-function studies of IRGQ without the complexity of monoclonal selection, offering a heterogeneous pool of edited cells suitable for pooled functional assays.
The HAP1 cell line is a chronic myeloid leukemia (CML)-derived, BCR-ABL-positive near-haploid line established as a robust platform for CRISPR-based functional genomics. Its near-haploid karyotype simplifies gene editing and reduces genetic redundancy, while retaining key signaling pathways, including interferon-gamma (IFN??) responsiveness, making it an ideal host for dissecting pathways related to autophagy and innate immunity.
IRGQ, a member of the immunity-related GTPase (IRG) family, is central to autophagy-dependent innate immunity. Its expression is induced by interferon-gamma (IFN??) via JAK-STAT signaling, with STAT1 and IRF1 acting as key transcriptional activators. IRGQ localizes to autophagosomal membranes, interacting with ATG5 and ATG12 to promote LC3-I lipidation to LC3-II, driving autophagosome maturation and lysosomal degradation. This process is critical for clearing ubiquitinated cargo, including p62/SQSTM1, and intracellular pathogens. Knockout of IRGQ blocks autophagic flux, causing LC3-II and p62 accumulation, and disrupts pathogen clearance.
In the HAP1 background, the IRGQ knockout model provides a physiologically relevant system to study the intersection of IFN?? signaling and autophagy. HAP1 cells maintain intact upstream signaling through IFNGR, JAK1, and STAT1, permitting direct investigation of how IRGQ mediates effector functions downstream of cytokine activation. This model is particularly valuable for elucidating the molecular mechanisms by which IRGQ coordinates autophagosome formation and cargo recognition, and for distinguishing IRGQ-dependent from IRGM- or other IRG-mediated processes. The polyclonal nature of the population avoids the clonal selection artifacts that can arise in single-cell clones, allowing for a more representative assessment of gene function across a variety of genetic backgrounds within the edited pool.
Applications of these IRGQ knockout cells span a broad range of experimental workflows in autophagy and innate immunity research. They are ideally suited for western blotting analyses of LC3-II and p62 levels to monitor autophagic flux under basal and IFN??-stimulated conditions. Immunofluorescence microscopy can be employed to visualize LC3 puncta formation and autophagosome accumulation. Co-immunoprecipitation experiments enable mapping of IRGQ??s interactions with ATG5, ATG12, and other autophagy machinery. Pathogen infection assays with model intracellular bacteria or parasites, such as Salmonella typhimurium or Toxoplasma gondii, provide a direct readout of IRGQ??s role in host defense. Furthermore, the cells facilitate functional genomics screens, autophagy-modulating drug discovery, and mechanistic studies of IRGQ-associated signaling networks. For additional product information or technical support, please contact Ascent Research.
