The DOK4 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for gene disruption of the DOK4 locus in the human HAP1 cell line. This loss-of-function model enables investigation of DOK4-dependent signaling processes without perturbing the broader genetic landscape, offering a consistent and scalable tool for functional studies.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) clone, characterized by a stable near-haploid karyotype that simplifies genetic manipulation and reduces functional redundancy from a second allele. Lacking functional p53, these cells are permissive for genome editing and are widely employed in functional genomics screens, making them an ideal host for generating knockout models that require unambiguous genotype-phenotype correlation.
DOK4 functions as an adaptor protein downstream of receptor tyrosine kinases, including the insulin receptor (INSR) and the RET receptor. Upon ligand stimulation, DOK4 becomes tyrosine-phosphorylated and recruits signaling proteins such as RASA1 (p120 RasGAP), NCK1, and CRK. This recruitment modulates key intracellular pathways, notably the Ras-MAPK cascade (via GRB2/SOS/HRAS/RAF1/MEK/ERK) and the PI3K-AKT pathway, thereby regulating cellular proliferation, differentiation, and metabolic responses as evidenced in insulin signaling and neurotrophin signaling contexts.
In the HAP1 background, DOK4 disruption eliminates the adaptor??s scaffolding function, allowing researchers to dissect its specific contributions to receptor tyrosine kinase signaling. The near-haploid nature ensures that each cell carries only one disrupted allele, simplifying interpretation of downstream effects. This model is particularly relevant for studying insulin resistance and type 2 diabetes mechanisms, as DOK4 mediates INSR-dependent metabolic signaling. Additionally, its role in RET-mediated neurodevelopment and oncogenesis provides a platform for exploring neuroblastoma and related pathologies in a genetically clean background.
Typical applications include functional genomics screens to identify modifiers of insulin sensitivity, quantitative assessment of signaling pathway activation via phospho-AKT and phospho-ERK assays, co-immunoprecipitation to map interactions with INSR or RET, and drug target validation studies. The polyclonal cell population is well-suited for standard assays such as Western blotting, RT-qPCR, immunofluorescence, and flow cytometry. For detailed technical specifications and ordering information, please contact Ascent Research.
