The CD248 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for functional investigation of CD248 (endosialin) in a near-haploid human background. This product provides a genetically disrupted CD248 locus across a heterogenous pool of HAP1 cells, enabling robust loss-of-function studies without clonal selection. The polyclonal nature avoids clone-specific artifacts while preserving the convenience of a stable knockout model for biochemical and cell-based assays.
The HAP1 cell line originates from the chronic myeloid leukemia-derived KBM-7 line and retains a fibroblastoid, adherent morphology with a near-haploid karyotype of approximately 25 chromosomes. This simplified genetic landscape reduces functional redundancy and enhances the utility of the line for genetic screens, signaling pathway dissection, and drug-target validation. HAP1 cells are widely used in biomedical research due to their ease of culture and compatibility with high-throughput applications.
CD248 is a type I transmembrane glycoprotein that functions as a receptor for extracellular matrix components, including fibronectin and collagen type I. Engagement of CD248 promotes integrin ??2??1 clustering and activates focal adhesion kinase (FAK) and Src family kinases, leading to downstream phosphorylation of ERK/MAPK and AKT. These cascades transcriptionally upregulate matrix metalloproteinases such as MMP-2 and MMP-9, fostering pericellular proteolysis and cell migration. Upstream, CD248 expression is induced by TGF-??1, HIF-1??, Wnt3a, and PDGF-BB, situating it at the intersection of fibrotic, hypoxic, and angiogenic signaling networks.
In the HAP1 fibroblastoid context, CD248 disruption provides a physiologically relevant platform to study stromal contributions to tumor progression and tissue remodeling. The near-haploid genome minimizes off-target effects and facilitates straightforward interpretation of phospho-signaling changes measured by immunoblotting. As HAP1 cells exhibit constitutive adhesion and migration properties, the knockout population is particularly suited for dissecting integrin-mediated mechanotransduction without the confounding influence of aneuploidy-driven signaling variability.
Applications include western blotting for CD248 protein verification, RT-qPCR for transcript confirmation, and immunofluorescence to assess localization changes upon gene disruption. Functional assays such as scratch-wound migration, cell adhesion to fibronectin- or collagen-coated surfaces, and collagen gel contraction can quantify the contribution of CD248 to matrix remodeling. Co-culture angiogenesis assays and gelatin zymography for MMP activity further extend the model to vascular biology. Phospho-FAK immunoblotting and flow cytometry for surface integrins allow detailed pathway analysis. For additional technical information or to explore custom assay configurations, please contact Ascent Research.
