The CASP5 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 line, with disruption of the CASP5 gene to ablate caspase-5 expression. This heterogeneous pool enables loss-of-function studies while maintaining population-level diversity, offering a robust system for investigating caspase-5-dependent processes in a near-haploid background, including inflammasome signaling, cytokine maturation, and pyroptosis.
HAP1 is a near-haploid fibroblast-like cell line derived from the chronic myeloid leukemia line KBM-7, originally from a male patient. Its haploid karyotype??with a single copy of most chromosomes except a disomic region of chromosome 15??facilitates genetic manipulation and phenotypic interpretation, making it ideal for functional genomics and haploid screens. HAP1 cells retain innate immune and cell death signaling pathways and offer adherent growth with a stable karyotype for reproducible long-term experiments.
Caspase-5 (CASP5) is an inflammatory caspase that mediates innate immune responses. It is recruited to inflammasome complexes formed by sensors including NLRP3, NLRC4, and AIM2, in conjunction with the adaptor PYCARD (ASC). Upon activation, CASP5 cleaves pro-IL-1?? and pro-IL-18 into bioactive cytokines and processes gasdermin D (GSDMD) to execute pyroptosis. Upstream, Toll-like receptor ligands, TNF, and IFN-?? prime inflammasome responses, while downstream, IL-1?? and IL-18 engage receptors to amplify NF-??B-dependent transcription. CASP5 interacts with PYCARD and cooperates with caspase-1 to link pathogen- and danger-associated molecular patterns to inflammatory cell death.
In the HAP1 background, CASP5 disruption provides a streamlined genetic system to dissect inflammasome pathways without diploid redundancy. The near-haploid genotype ensures loss-of-function phenotypes are not masked, enabling clear attribution of defects. This model is advantageous for synthetic lethality screens, CRISPR perturbations, and epistasis analyses in innate immunity and cell death. Additionally, HAP1??s incomplete interferon pathway reduces type I interferon crosstalk, simplifying cytoplasmic inflammasome activation studies. Researchers can thus precisely characterize CASP5 in responses to cytosolic pathogens, metabolic stress, or pharmacological modulators.
These CASP5 knockout HAP1 cells enable diverse inflammasome research applications, including western blotting for caspase-5, cleaved IL-1??, and GSDMD; ELISA for IL-1?? and IL-18; LDH release assays for pyroptosis; co-immunoprecipitation for caspase-5 interactions; immunofluorescence for ASC specks; RT-qPCR for inflammasome components; and flow cytometry for cell death. They support studies in inflammatory disorders, autoimmunity, cancer immunity, and sepsis, providing a reliable platform for inhibitor testing and pathway validation. For further technical information or custom experimental inquiries, please contact Ascent Research.
