The CD2AP Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 cell line, featuring disruption of the CD2AP gene. This loss-of-function model enables investigation of CD2AP??s scaffold function in linking membrane receptors to the actin cytoskeleton, without requiring single-cell cloning. The heterogeneous knockout population provides a reproducible system for studying endocytosis, adhesion, and signal transduction.
HAP1 is a near-haploid human cell line from a chronic myelogenous leukemia patient (KBM-7), adherent, with wild-type TP53 and myeloid lineage features. Its haploid genome simplifies gene targeting and reduces genetic redundancy, making it ideal for high-throughput CRISPR screens and unambiguous genotype-phenotype analysis in hematopoietic cancer research.
CD2AP is an adaptor protein that connects membrane receptors such as CD2 and nephrin to the actin cytoskeleton. It is activated by T-cell receptor engagement and integrin signaling, and it regulates downstream effectors including WASp, the Arp2/3 complex, Rac1, and Cdc42 to promote actin polymerization. In podocytes, CD2AP complexes with nephrin and podocin to maintain the slit diaphragm, while its interactions with cortactin, CIN85, CAPZA1, and SHIP2 modulate endocytosis and focal adhesion dynamics. CD2AP also transmits signals via the PI3K-AKT pathway, governing cell survival and migration.
In HAP1 cells, CD2AP knockout disrupts actin-based processes critical for leukemic cell migration and invasion. The near-haploid background allows reconstitution of signaling pathways by expressing exogenous receptors, enabling studies of CD2AP-dependent T-cell activation or podocyte filtration in a tractable cell model. This polyclonal knockout population is particularly suited for dissecting CD2AP??s role in diseases like focal segmental glomerulosclerosis and Alzheimer disease, where CD2AP mutations impair cytoskeletal and endocytic functions.
Researchers can use this product for western blotting and RT-qPCR to confirm CD2AP loss, co-immunoprecipitation to examine interactions with nephrin or CD2, and immunofluorescence to visualize actin stress fibers. Transwell migration and endocytosis assays quantify functional defects. Flow cytometry for CD2 surface expression complements biochemical analyses. Applications span podocyte biology, T-cell signaling, Alzheimer research, and cancer metastasis studies. For further details, contact Ascent Research.
