The CD1D Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the human CD1D gene in the HAP1 near-haploid cell line. This product provides a heterogeneous pool of cells carrying targeted gene disruptions, enabling loss-of-function studies of CD1D in a genetically tractable background. The polyclonal format captures diverse mutational events across the population, facilitating robust phenotypic analysis without the clonal selection biases inherent in single-cell-derived knockouts.
HAP1 cells are a human near-haploid cell line originally derived from the KBM-7 chronic myeloid leukemia cell line. Their near-haploid karyotype simplifies genetic manipulation and reduces functional redundancy from diploid gene copies, making them an ideal host for knockout-based genetic screens and mechanistic studies. The HAP1 line has been widely adopted as a model system for investigating gene function in pathways related to cancer biology, immunology, and cell signaling.
CD1D encodes a non-classical major histocompatibility complex class I-like glycoprotein that presents lipid antigens to invariant natural killer T (iNKT) cells. CD1D forms heterodimers with beta-2-microglobulin and loads lipid antigens in endosomal compartments for presentation to the iNKT T-cell receptor. This recognition triggers rapid secretion of cytokines such as IFN-gamma and IL-4, thereby bridging innate and adaptive immunity. Upstream regulators include IL-4, IFN-gamma, Toll-like receptor ligands, and microbial lipid antigens, while downstream effector responses involve iNKT cell activation, IFN-gamma and IL-4 production, and dendritic cell maturation. The CD1D pathway engages key interacting factors such as beta-2-microglobulin, lipid antigens, iNKT TCR, and CD1d-restricted T cells, and converges on signaling components including SLAM family receptors.
Disruption of CD1D expression in HAP1 cells offers a powerful model to dissect the molecular requirements for lipid antigen presentation and iNKT cell activation. The near-haploid genetic background of HAP1 ensures that even small perturbations in CD1D function yield clear phenotypes, facilitating quantitative analysis of immune synapse formation, cytokine profiles, and downstream signaling. By eliminating CD1D-mediated antigen presentation, this knockout population allows researchers to isolate the contributions of CD1D-dependent iNKT cell responses from other innate and adaptive pathways.
These cells are well-suited for a broad range of immunological and cell biological applications. They can be employed in flow cytometry-based assays to assess iNKT cell activation or in ELISA-based cytokine secretion studies to measure IFN-gamma and IL-4 release upon lipid antigen stimulation. Lipid antigen presentation assays, iNKT cell proliferation co-culture experiments, and immunofluorescence microscopy to examine CD1D subcellular localization are also directly applicable. Further, the cells support western blotting for CD1D expression analysis and functional reconstitution studies with CD1D variants. These tools are valuable in research areas including autoimmune diseases, infections, cancer, and inflammatory diseases. For additional technical information about these polyclonal knockout cells, please contact Ascent Research.
