The EFCAB14 Knockout Jurkat Polyclonal Cells provide a polyclonal pool of CRISPR/Cas9-edited Jurkat T cells with targeted disruption of the EFCAB14 gene. This loss-of-function model is designed for investigating the role of the putative EF-hand calcium-binding protein EFCAB14 in T-cell signaling. The polyclonal format ensures a diverse population of knockout cells, suitable for studying gene function without clonal selection artifacts.
The parental Jurkat cell line is derived from a 14-year-old male with acute T-cell leukemia and serves as a model for T lymphocyte activation, signaling, and apoptosis. Widely used in TCR signaling research, Jurkat cells offer well-defined pathways for calcium mobilization and cytokine production. Their established use in CRISPR/Cas9 editing makes them an optimal host for gene disruption studies.
EFCAB14 encodes an EF-hand domain-containing protein suggesting a role in calcium-dependent signal transduction. In Jurkat T cells, TCR stimulation activates ZAP70, LAT, and PLC??1, triggering IP3-mediated calcium release. Elevated calcium activates calcineurin, which dephosphorylates NFAT transcription factors. EFCAB14 may interact with calmodulin and other EF-hand proteins to modulate this pathway. Knockout of EFCAB14 is predicted to alter calcium homeostasis, impairing calcineurin/NFAT and CAMK kinase signaling, and disrupting cytokine expression and activation-induced apoptosis.
Loss of EFCAB14 in Jurkat cells is expected to impact calcium flux, NFAT-driven transcription, and T-cell activation. This model is relevant for studying T-cell leukemia, autoimmune disorders, and immunodeficiencies linked to aberrant calcium signaling. By assessing proliferation, IL-2 secretion, and apoptosis, researchers can delineate EFCAB14’s role in immune regulation.
This polyclonal knockout pool enables a range of assays, including Fluo-4 calcium flux, NFAT luciferase reporter, IL-2 ELISA, phospho-ZAP70 western blot, CD69 flow cytometry, and Annexin V apoptosis detection. RNA-seq can characterize transcriptomic changes. Applications include functional genomics, immune modulator screening, and signaling pathway analysis. For further information, contact Ascent Research.
