The DOCK8 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population engineered to disrupt the DOCK8 gene in the human HAP1 cell line. This loss-of-function model exploits the near-haploid background of HAP1 cells, enabling straightforward functional analysis of DOCK8. The heterogeneous pool of edited cells minimizes clonal artifacts and is well-suited for population-based assays in functional genomics, signaling, and phenotypic screening.
HAP1 cells are a human near-haploid chronic myeloid leukemia line derived from KBM-7, exhibiting hematopoietic progenitor features and suspension growth. Their haploid karyotype simplifies genetic disruption and phenotypic readouts, as single-allele knockout can uncover phenotypes. HAP1 cells maintain key signaling cascades of hematopoietic lineages, making them a versatile platform for studying immune cell biology, protein interactions, and drug responses.
DOCK8 functions as a guanine nucleotide exchange factor specifically activating Cdc42 and Rac1, thereby regulating actin cytoskeleton reorganization, cell migration, and immune cell activation. Upstream activation occurs through the TCR/CD3 complex, B-cell receptor, IL-2 and IL-4 receptors, and the chemokine receptor CXCR4, mediated by PI3K and Src family kinases. DOCK8 then interacts with WASP, cortactin, and CrkL to stimulate Cdc42/Rac1 signaling, leading to PAK kinase activation, WASP/WAVE complex recruitment, and ARP2/3-mediated actin polymerization. This signaling cascade ultimately promotes lymphocyte migration, immunological synapse formation, and connects to NF-??B and MAPK transcriptional programs.
Within HAP1 cells, DOCK8 knockout disrupts Cdc42/Rac1-dependent pathways, providing a surrogate model mirroring lymphocyte dysfunction observed in autosomal recessive hyper-IgE syndrome and combined immunodeficiency. The hematopoietic origin of HAP1 cells ensures endogenous expression of relevant adaptors such as WASP, cortactin, and ELMO1, enabling dissection of actin dynamics and migration defects. This model facilitates investigation of pathogenic mechanisms and identification of therapeutic targets for primary immunodeficiencies.
This DOCK8 knockout product supports diverse applications including functional genomics, CRISPR-based genetic interaction screens, and drug discovery efforts aimed at hyper-IgE syndrome and other immunodeficiencies. Researchers can assess DOCK8 protein levels via Western blot, transcript expression by RT-qPCR, immune marker expression by flow cytometry, and cell migration using transwell assays. Additionally, actin polymerization assays, Cdc42/Rac activation measurements, and RNA sequencing allow comprehensive network analysis. These polyclonal knockout cells offer a robust and scalable resource for mechanistic studies and high-throughput screening. For additional product details, please contact Ascent Research.
