The CD160 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout population carrying targeted disruptions in the CD160 gene. This pool of gene-edited HAP1 cells provides a heterogeneous yet functionally relevant model for studying CD160 loss-of-function, avoiding clonal selection biases. The polyclonal format is ideal for applications requiring large-scale genetic perturbation, such as pooled screening and population-level functional assays.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) isolate, exhibiting a fibroblastoid morphology. With a haploid karyotype (except for parts of chromosomes 8 and 15), HAP1 offers a simplified genetic system that enhances the efficiency of CRISPR-mediated knockout generation and phenotype analysis, making it a widely used platform for gene-function studies in a myeloid lineage context.
CD160 encodes a glycosylphosphatidylinositol (GPI)-anchored receptor predominantly expressed on natural killer (NK) cells and T cell subsets. It functions as an immune checkpoint molecule by binding the herpes virus entry mediator (HVEM; TNFRSF14) and major histocompatibility complex class I (HLA-A/B/C) ligands. HVEM engagement triggers downstream signaling through NF-??B, PI3K/AKT, and ERK pathways, leading to the regulation of interferon-?? production and cytolytic activity. The signaling network is further modulated by competitive interactions with LIGHT (TNFSF14) and BTLA. CD160 expression is inducible by cytokines IL-15 and IL-2, T cell receptor stimulation, and viral infection.
In the HAP1 background, the single-copy nature of most genes, including CD160, ensures that CRISPR-mediated gene disruption leads to complete loss of protein function across the polyclonal pool. This near-haploid model thus provides a highly penetrant knockout system for investigating CD160-mediated immune checkpoint pathways despite the non-hematopoietic origin of HAP1 cells. When combined with ectopic expression of NK or T cell co-receptors, these knockout cells allow reconstitution of CD160-HVEM signaling modules, enabling detailed dissection of signal transduction, ligand-receptor specificity, and downstream transcriptional responses in a clean genetic environment.
Applications include immuno-oncology research, autoimmune disease modeling, and studies of host?Cpathogen interactions where CD160 plays a modulatory role. Representative assays such as flow cytometry for HVEM binding, NK cell cytotoxicity (chromium release) with HAP1 as target cells, ELISA for IFN-??, western blotting for phospho-AKT, and RNA-seq-based transcriptomics are readily supported. The knockout cells serve as an isogenic control for CD160-dependent phenotypes and facilitate high-throughput screening for checkpoint inhibitors. For further information, please contact Ascent Research.
