This product is a polyclonal CRISPR/Cas9-edited HAP1 cell population with targeted disruption of the IFNGR1 gene, creating a loss-of-function model for interferon gamma receptor signaling. The polyclonal format generates a heterogeneous pool of edited alleles, suitable for bulk functional assays and pooled screening approaches where population-level responses are informative. This knockout tool enables systematic dissection of IFN-??-mediated pathways without the need for clonal isolation.
HAP1 cells are a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia background. Their haploid karyotype simplifies genome engineering and facilitates direct genotype?Cphenotype correlations, establishing them as a premier model for CRISPR-based functional genomics, knockout screens, and signal transduction studies. Despite their malignant origin, HAP1 cells retain intact core signaling modules responsive to cytokines, including interferon gamma.
IFNGR1 encodes the ligand-binding alpha chain of the interferon gamma receptor. Upon IFN-?? binding, it heterodimerizes with IFNGR2, activating the associated kinases JAK1 and JAK2. These phosphorylate STAT1, which homodimerizes and translocates to the nucleus to drive expression of immune effector genes such as IRF1, CIITA, CXCL10, and MHC class I/II. Negative regulation is exerted by SOCS1 and the phosphatase PTPN2, while upstream cytokines like IL-12 and IL-18 promote IFN-?? production in immune settings.
Disruption of IFNGR1 in the haploid HAP1 background eliminates responsiveness to IFN-??, as evidenced by loss of STAT1 phosphorylation, IRF1 induction, and surface MHC class I upregulation. This genetic ablation provides a clean platform for epistatically probing JAK-STAT components, reconstituting mutant receptors, or performing suppressor and enhancer screens. The simplified genetic landscape of HAP1 cells ensures that knockout phenotypes are not confounded by multiple allelic copies.
Researchers can leverage these cells to dissect IFN-?? signaling in infectious disease, autoimmunity, and cancer immunology. Typical experimental approaches include interferon gamma stimulation combined with Western blot analysis of phospho-STAT1, RT-qPCR quantitation of IRF1 and CIITA transcripts, and flow cytometric measurement of MHC class I surface expression. The polyclonal population is also amenable to genome-wide CRISPR modifier screens for factors that restore or modulate IFN-?? sensitivity. For additional product details or custom editing services, please contact Ascent Research.
