The CD226 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population in the near-haploid human HAP1 cell line. This product features a heterogeneous pool of CD226-deficient cells, providing a robust loss-of-function model for functional studies without clonal selection. The polyclonal format maintains population diversity while achieving effective disruption of the CD226 gene, making it suitable for assays where averaged population responses are informative, including high-throughput screening and signaling pathway analyses.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid karyotype reduces genetic redundancy, facilitating efficient gene targeting and straightforward genotype interpretation. As a neoplastic hematopoietic cell line, HAP1 expresses relevant immunoreceptors and signaling molecules, making it an excellent host for studying genes involved in immune cell adhesion, activation, and cytotoxicity. The CD226 knockout in this platform allows direct investigation of immunoreceptor functions in a simplified yet physiologically meaningful context.
CD226 (DNAM-1) is an immunoreceptor that binds CD155 and CD112, promoting immune synapse formation and downstream signaling crucial for T cell and NK cell effector functions. Ligand engagement activates Fyn kinase, which triggers the PI3K/AKT and MAPK/ERK pathways, leading to LFA-1 activation, actin polymerization, and enhanced cytokine production. Upstream regulators include TCR engagement, IL-2, and IL-15, which modulate CD226 expression and activity. This signaling integrates adhesion with co-stimulatory signals, making CD226 a key mediator in immune surveillance and autoimmune regulation.
Disrupting CD226 in HAP1 cells ablates ligand-induced activation of PI3K/AKT and MAPK/ERK pathways, providing a clear model to dissect CD226-proximal signaling events. The hematopoietic origin and expression of relevant pathway components such as Fyn, LFA-1, and integrins enable biochemical and functional analyses of adhesion-dependent signaling. This knockout model is particularly valuable for studying immune checkpoint regulation and the interplay between CD226 and inhibitory receptors competing for CD155 and CD112. The near-haploid background minimizes confounding allelic effects, facilitating clean phenotypic and signaling readouts.
This product is ideal for cancer immunotherapy, autoimmune disease modeling, NK cell biology, and T cell activation research. Researchers can employ flow cytometry to confirm CD226 loss, western blotting for Fyn and phospho-AKT analysis, cytotoxicity and cell adhesion assays to evaluate functional impairment, and cytokine ELISAs to measure altered interferon-?? secretion. It also supports drug target screening and CRISPR modifier screens to identify pathway dependencies. For further inquiries, please contact Ascent Research.
