The IBTK Knockout Jurkat Polyclonal Cells product comprises a polyclonal population of Jurkat T lymphocytes modified by CRISPR/Cas9-mediated disruption of the IBTK gene. This approach generates a heterogeneous pool of cells bearing various loss-of-function alterations at the target locus, which collectively ablate IBTK protein expression. The polyclonal format recapitulates the diverse genetic outcomes of genome editing and provides a robust, reproducible model for studying IBTK function without the selection bias inherent in single-cell clones. These cells are ideally suited for high-throughput screening and biochemical assays requiring a renewable knockout resource.
Jurkat cells are an immortalized T lymphocyte line from an acute T cell leukemia patient, serving as a standard model for T-cell activation and apoptosis. They express the TCR/CD3 complex and respond to stimulation with rapid phosphorylation events, calcium mobilization, and cytokine production. The cell line has been pivotal in elucidating TCR-proximal signaling, including the roles of kinases LCK, ZAP70, ITK and adaptors LAT, SLP76. Its well-mapped signaling network and ease of genetic manipulation make it an excellent host for studying regulatory proteins such as IBTK.
IBTK encodes an inhibitor of Bruton’s tyrosine kinase (BTK) and IL-2-inducible T-cell kinase (ITK), serving as a negative regulator of TCR signaling. Upon TCR stimulation, IBTK interacts with BTK and attenuates its kinase activity, reducing phospholipase C gamma 1 (PLC??1) phosphorylation and inositol trisphosphate production. This limits calcium mobilization and dampens calcineurin/NFAT and NF-??B activation, pathways essential for T-cell proliferation and cytokine synthesis. IBTK also binds actin and the adaptor PINCH1, thereby influencing actin cytoskeleton dynamics critical for immune synapse function.
Disruption of IBTK in Jurkat cells removes a key inhibitory constraint on TCR signaling, resulting in heightened calcium responses, increased NFAT and NF-??B transcriptional activity, and elevated IL-2 secretion. This hyperactivation phenotype is particularly relevant for modeling T-cell leukemogenesis, where constitutive activation of these pathways drives malignant proliferation. The polyclonal knockout population exhibits a consistent functional phenotype across a diverse allelic spectrum, confirming that IBTK loss is the primary determinant. Moreover, the Jurkat background provides a simplified, tractable system to study the interplay between BTK/ITK-mediated signals and actin dynamics in T cells.
Researchers can utilize these cells in a variety of assays, including Western blotting for phosphorylated BTK, ITK, PLC??1, and NF-??B; flow cytometry to measure intracellular IL-2 production or calcium flux; and reporter assays for NFAT and NF-??B activity. The model is also suitable for co-immunoprecipitation studies to map IBTK interaction networks and for apoptosis assays employing annexin V or caspase-3 cleavage. The polyclonal nature makes the cells particularly advantageous for pharmacological inhibitor screens targeting BTK/ITK, as the heterogeneous target population mimics the genetic variability that may be encountered in clinical settings. For further information or to request a quotation, please contact Ascent Research.
