The CD58 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for the targeted disruption of the CD58 gene in the near-haploid HAP1 cell line. This product enables loss-of-function studies of CD58, a key adhesion molecule involved in T cell?Cantigen-presenting cell (APC) interactions. The polyclonal format ensures a heterogeneous pool of edited cells, facilitating robust and reproducible genetic screens and functional assays without the need for single-cell cloning.
HAP1 is a human near-haploid cell line derived from the KBM-7 chronic myeloid leukemia (CML) line and lacks a functional p53 tumor suppressor pathway. Its haploid karyotype simplifies gene editing and genotypic analysis, as disruptions in single alleles result in clear loss-of-function phenotypes. The hematopoietic origin of HAP1 cells makes them particularly suitable for immunological and hematological disease modeling, providing a relevant cellular context for studying CD58-mediated pathways in a malignancy-derived background.
CD58 (LFA-3) functions as a ligand for CD2, mediating the adhesion between T cells and APCs that is essential for immunological synapse formation and costimulatory signaling. In T cells, CD58 engagement activates downstream kinases including LCK and ZAP70, which propagate signals through the PI3K/AKT and MAPK cascades, ultimately regulating cytokine production such as IL-2. CD58 expression is transcriptionally controlled by pro-inflammatory cytokines and transcription factors, including TNF-??, IFN-??, IL-1??, and NF-??B. Additionally, CD58 interacts with the actin cytoskeleton via filamin A, linking adhesion to cytoskeletal reorganization.
Disruption of CD58 in HAP1 cells abolishes the CD2-CD58 interaction, impairing the initiation of T cell receptor-dependent signaling and costimulatory pathways. In the haploid HAP1 background, the polyclonal CD58 knockout population enables straightforward genotype-phenotype correlations, as each cell harbors a disrupted CD58 allele. This model is particularly valuable for dissecting the molecular requirements of immune synapse assembly, T cell activation thresholds, and adhesion-dependent signaling in a high-throughput-compatible format, avoiding the complexity of diploid genome compensation.
Researchers can employ these CD58 knockout HAP1 polyclonal cells in a variety of experimental workflows, including flow cytometry to confirm loss of CD58 surface expression, co-culture assays with T cells to measure adhesion defects, phospho-flow cytometry to assess signaling activation (e.g., phosphorylated LCK, AKT, or ERK), and RT-qPCR for downstream cytokine gene transcription. This product is well-suited for functional genomic screens, drug target validation efforts, and mechanistic studies in autoimmunity and graft-versus-host disease. For additional details and technical assistance, please contact Ascent Research.
