The CCDC47 Knockout Jurkat Polyclonal Cells comprise a heterogeneous population of CRISPR/Cas9-edited Jurkat T-lymphoblastoid cells harboring targeted disruption of the CCDC47 gene locus. This polyclonal knockout model enables loss-of-function investigations in an immortalized human T-cell background, avoiding clonal selection biases and preserving a distribution of genetic disruptions across the population. The edited cells are provided as a ready-to-use reagent for studying endoplasmic reticulum (ER) homeostasis and unfolded protein response (UPR) pathways in a lymphoid context, without requiring single-cell isolation or characterization of individual clones. Researchers can employ these polyclonal knockout cells directly in functional assays, facilitating rapid screening and mechanistic dissection of CCDC47-dependent processes.
The host Jurkat cell line is a widely used leukemic T-lymphoblast model originally derived from an acute T-cell leukemia patient. Jurkat cells recapitulate key aspects of T-cell receptor signaling, apoptosis, and immunological activation, making them an ideal platform for examining how ER stress intersects with T-cell biology. Their robust growth in suspension culture and well-characterized signaling pathways offer a tractable system for genetic perturbation studies, particularly in the context of calcium mobilization and stress-induced cell death that are central to lymphocyte function.
CCDC47 encodes an ER-resident transmembrane protein that serves as a cofactor for the sarcoplasmic/endoplasmic reticulum calcium ATPase SERCA2 (ATP2A2) and interacts directly with the lectin chaperone calnexin (CANX). Through these associations, CCDC47 modulates ER calcium sequestration and facilitates oxidative folding of calnexin client proteins. Disruption of CCDC47 leads to compromised calcium uptake, accumulation of misfolded proteins, and consequent activation of the UPR. The UPR is orchestrated by three canonical sensors: IRE1 (ERN1), PERK (EIF2AK3), and ATF6. Downstream transcriptional effectors include XBP1, ATF4, and CHOP (DDIT3), which together with the master chaperone BiP (HSPA5) govern adaptive and pro-apoptotic responses. CCDC47 thus integrates calcium and protein folding quality control, with its loss triggering a cascade through these UPR components.
In Jurkat T lymphocytes, CCDC47 ablation provides a unique model for investigating how ER proteostatic perturbations influence T-cell physiology. Jurkat cells rely on precise calcium oscillations for signaling and apoptosis, and interruption of SERCA2-mediated calcium reuptake will sensitize these pathways. The knockout cells allow dissection of UPR-dependent modulation of T-cell receptor downstream events, apoptosis sensitivity, and cytokine response, with direct relevance to T-cell acute lymphoblastic leukemia and congenital disorders of glycosylation. Moreover, since Jurkat cells express the neurodevelopmental disease-associated splice variants, this model may illuminate CCDC47??s role in nervous system pathologies, bridging hematological and neurological research.
These polyclonal CCDC47 knockout cells are suitable for a spectrum of applications including quantitative assessment of UPR activation by western blotting for BiP, CHOP, and phospho-eIF2??; monitoring of calcium flux with fluorescent indicators such as Fluo-4; and flow cytometric analysis of apoptosis via annexin V staining. RT-qPCR can quantify XBP1 splicing efficiency, while co-immunoprecipitation experiments enable mapping of calnexin and PDI interactions. Immunofluorescence microscopy reveals ER morphological changes, and these cells can serve as a background for chemical or genetic screens targeting ER stress modulators. For further information or technical inquiries, please contact Ascent Research.
