The ALCAM Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HAP1 cells with targeted disruption of the human ALCAM gene, encoding activated leukocyte cell adhesion molecule (CD166). Supplied as a mixed polyclonal pool, this product captures diverse editing events without clonal selection artifacts. The CRISPR/Cas9-mediated gene disruption yields a loss-of-function allele, abrogating ALCAM protein expression and its biological activities. This knockout model offers a powerful in vitro system for studying ALCAM-mediated processes in a genetically defined, haploid context.
HAP1 is a near-haploid, adherent human cell line derived from the male KBM-7 chronic myeloid leukemia (CML) line. Its haploid karyotype for most chromosomes simplifies genetic analysis, enabling direct phenotype?Cgenotype correlations. HAP1 cells are widely used in functional genomics, genetic screens, and gene-function studies due to their stable near-haploid state, which ensures that single-gene disruptions produce complete null phenotypes unconfounded by second alleles.
ALCAM (CD166) is a transmembrane immunoglobulin superfamily protein mediating homophilic (ALCAM?CALCAM) and heterophilic (ALCAM?CCD6) cell adhesion. Homophilic binding maintains tissue architecture and tumor cell clustering, while CD6 engagement on T cells promotes T-cell activation and immunological synapse formation. ALCAM transcription is induced by TNF-?? and IL-1?? through NF-??B and AP-1. Downstream, ALCAM signals via RhoA, Rac1, ERK1/2, and Akt, leading to F-actin reorganization and modulation of cell motility and invasion. Additionally, ALCAM interacts with L1CAM and NgR1, implicating it in neurite outgrowth and axon guidance. Consequently, ALCAM disruption impairs both cell?Ccell adhesion and multiple intracellular signaling cascades.
In the HAP1 haploid background, ALCAM knockout provides an unambiguous loss-of-function model, as the single gene disruption directly yields a null phenotype without allelic compensation. This cellular system is well-suited for quantifying adhesion, migration, and invasion via standard assays, and for imaging actin cytoskeleton dynamics. Complete absence of surface CD166 is verifiable by flow cytometry, while RT-qPCR and Western blot confirm transcript and protein depletion. Co-immunoprecipitation can assess disrupted ALCAM interactions with CD6, L1CAM, or NgR1. The polyclonal nature ensures robustness against clonal variability.
These polyclonal ALCAM knockout cells are suited for studies of cancer metastasis in melanoma, prostate, breast, and colorectal carcinomas, as well as for investigating T-cell costimulation and immune synapse dynamics. They also serve as a platform for neurodevelopmental research examining axon guidance, and for high-throughput drug screens targeting ALCAM-dependent signaling. In inflammatory disease models, including rheumatoid arthritis and multiple sclerosis, they facilitate analysis of leukocyte transendothelial migration. For further technical details or to request a quotation, please contact Ascent Research.
