CD48 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa cervical adenocarcinoma cell line, engineered to disrupt the CD48 gene. This product provides a heterogeneous pool of cells harboring targeted loss-of-function mutations in the CD48 locus, enabling robust investigation of CD48-mediated signaling and adhesion in a human epithelial background. The polyclonal format preserves population-level gene disruption without single-cell cloning, offering a genetically diverse model for studying immune receptor-ligand interactions.
The parental HeLa cell line is an immortalized epithelial line originally derived from cervical adenocarcinoma, widely employed for its robust growth, high transfection efficiency, and extensively characterized signaling networks. As a foundational model in cancer biology, virology, and cell signaling, HeLa cells provide a reliable and reproducible host for CRISPR-based gene knockout studies. Their epithelial origin offers a distinct context for examining the functional consequences of CD48 loss, complementing traditional immune cell models.
CD48 encodes a glycosylphosphatidylinositol (GPI)-anchored cell-surface receptor that functions as a ligand for CD244 (2B4), a member of the signaling lymphocyte activation molecule (SLAM) family. CD48?CCD244 binding mediates bidirectional signaling that regulates immune cell adhesion, activation, and cytotoxicity. Downstream signal propagation involves the adaptor SH2D1A (SAP), Src kinase Fyn, phospholipase C-?? (PLC-??), and extracellular signal-regulated kinase (ERK). Expression of CD48 is transcriptionally controlled by cytokines such as interleukin-2 (IL-2) and interleukin-4 (IL-4) through the transcription factors NFAT and STAT5. The receptor also engages CD2 and SLAMF6, contributing to immunological synapse formation. In this knockout model, disruption of CD48 abolishes the CD48?CCD244 interaction, leading to impaired recruitment of SH2D1A and Fyn, reduced ERK and PLC-?? phosphorylation, and attenuation of downstream effector functions.
Although CD48 is predominantly expressed on hematopoietic cells, ectopic expression in HeLa cells permits dissection of CD48-dependent signaling modules in a well-defined epithelial system free from confounding immune-specific inputs. This model enables focused analysis of CD48?CCD244 binding and downstream signal transduction. Additionally, HeLa cells expressing CD48 can be used to investigate tumor?Cimmune crosstalk when co-cultured with CD244-expressing NK or T cells, isolating the contribution of CD48 to adhesion and signaling.
These CD48 knockout HeLa polyclonal cells are applicable to immune synapse studies, tumor immunology, and adhesion signaling research. They are suitable for co-culture cytotoxicity assays with CD244+ effector cells, enabling flow cytometric measurement of CD48-dependent killing. Loss of CD48 protein and mRNA can be confirmed by western blotting and RT-qPCR, while co-immunoprecipitation and proximity ligation assays quantify disrupted CD48?CCD244 complexes and adaptor recruitment. Phospho-specific antibodies can monitor ERK and PLC-?? activation status. For further information, please contact Ascent Research.
