The GSDMD Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting human GSDMD in the HAP1 near-haploid cell line. This product provides a loss-of-function model for studying pyroptosis and inflammasome-mediated cytokine release. Generated by CRISPR/Cas9-mediated gene disruption, the polyclonal population offers a heterogeneous pool of edited alleles appropriate for pooled functional studies without clonal selection.
HAP1 cells derive from the KBM-7 chronic myeloid leukemia line and exhibit a near-haploid karyotype, except for a disomic chromosome 8 region. This haploid genetic model enables efficient gene knockout studies since disruption of a single allele is sufficient for most loci. HAP1 cells are widely adopted for genetic screens and loss-of-function analyses of cell death and inflammatory pathways. Their simplified genome makes them particularly useful for dissecting complex processes like pyroptosis.
GSDMD is the executioner of pyroptosis. It is cleaved by inflammatory caspases??caspase-1, -4, and -5??upon activation of inflammasome sensors (NLRP3, NLRC4, AIM2, NLRP1) and the adaptor ASC (PYCARD). The released N-terminal fragment oligomerizes and forms plasma membrane pores, leading to cell lysis and release of IL-1??, IL-18, HMGB1, and LDH. Thus, GSDMD acts downstream of inflammasome assembly and upstream of cytokine secretion. This pathway is triggered by signals such as LPS, bacterial toxins, and DAMPs.
Knocking out GSDMD in HAP1 cells provides a clean genetic background to study inflammasome signaling. The near-haploid state ensures that phenotypes are directly attributable to GSDMD loss, eliminating complications from diploid gene redundancy. The polyclonal nature maintains diversity useful for pooled screening approaches, such as drug resistance profiling or modifier screens, while still enabling focused pathway analysis. This model allows clear dissection of GSDMD-dependent versus independent pyroptotic mechanisms.
Applications include mechanistic dissection of pyroptosis via Western blotting for GSDMD cleavage, LDH release assays, and IL-1??/IL-18 ELISA. These cells are suitable for live-cell imaging of pore formation, caspase activity assays, and screening of pyroptosis inhibitors. In drug discovery, they facilitate identification of compounds modulating inflammasome activity or blocking GSDMD pores. They also support research into autoinflammatory diseases, sepsis, neurodegeneration, and cancer where GSDMD-mediated inflammation plays a role. For further information, please contact Ascent Research.
