The CD24 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney epithelial cell line. In this product, targeted disruption of the CD24 gene generates a heterogeneous pool of knockout cells, each carrying distinct editing outcomes at the CD24 locus. This polyclonal format provides a loss-of-function model that circumvents clonal selection artifacts and reflects population-level gene ablation, making it a robust tool for interrogating CD24-dependent phenotypes without the bias of a single isogenic clone. The cells are supplied as a viable, proliferating culture ready for functional assays.
HEK293T cells are a widely adopted host in molecular and cellular biology due to their high transfection efficiency, robust protein expression capacity, and utility in viral packaging. Originally derived from HEK293 cells by stable integration of the SV40 large T antigen, the HEK293T line exhibits an adherent epithelial morphology and supports facile genetic manipulation. These features render HEK293T an ideal platform for studying exogenously introduced gene disruptions, enabling consistent and reproducible analysis of signaling pathways and cellular behaviors in an easily tractable epithelial background.
CD24 encodes a glycosylphosphatidylinositol (GPI)-anchored cell surface glycoprotein that mediates cell adhesion and intracellular signal transduction. CD24 resides within membrane lipid rafts, where it interacts with P-selectin (SELP), L1CAM, Siglec-10, and SRC-family kinases. Engagement of CD24 activates downstream cascades involving SRC, focal adhesion kinase PTK2 (FAK), and the PIK3CA?CAKT1 and MAPK1/MAPK3 (ERK1/2) axes, ultimately modulating transcription factors such as NFKB1 (NF-??B) and RELA, and promoting the expression of pro-invasive genes including MMP2, MMP9, and VEGFA. Upstream, CD24 expression is regulated by STAT3, ETS1, SP1, HIF-1A, and inflammatory cytokines IL6 and TNF. Knockout of CD24 therefore disrupts a nodal signaling hub that coordinates adhesion-dependent proliferation, migration, and survival signals.
In the HEK293T background, ablation of CD24 eliminates GPI-anchored signaling domains from the plasma membrane, dampening SRC/FAK-mediated adhesion signaling and attenuating both MAPK/ERK and PI3K/AKT pathway activity. This loss-of-function model recapitulates aspects of CD24 silencing observed in tumor and immune contexts, providing a tractable system to dissect CD24??s contribution to cell migration, invasion, and proliferation. Because HEK293T cells natively express low levels of endogenous CD24, the polyclonal knockout population offers a clean background for rescue experiments and for exploring CD24??s role independent of immune cell-specific co-receptors. Researchers can employ this model to examine how CD24 deficiency alters phospho-signaling dynamics, adhesion to P-selectin, and matrix degradation capabilities.
This polyclonal knockout cell product supports diverse applications, including detailed signaling studies using Western blotting for CD24, flow cytometric assessment of surface CD24 loss, and Transwell migration/invasion assays. Phospho-specific antibodies against AKT1 and ERK1/2 enable dissection of pathway alterations, while adhesion assays to recombinant P-selectin quantify CD24-dependent tethering. Proliferation assays and drug response experiments further delineate CD24??s involvement in growth and therapeutic resistance. Functional complementation by re-expression of CD24 or its signaling partners validates specific mechanisms. For detailed technical protocols and batch-specific knockout characterization, please contact Ascent Research.
