The C15orf40 Knockout Jurkat Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of Jurkat T lymphocytes harboring targeted disruption of the C15orf40 gene. This loss-of-function model enables systematic investigation of C15orf40-dependent regulatory mechanisms in a human immune cell context. The polyclonal format preserves heterogeneous editing events across the cell pool, reflecting a bulk knockout population without clonal selection.
The Jurkat cell line is an immortalized human T lymphocyte model originally derived from a patient with acute T cell leukemia. These suspension cells retain key features of T cell biology, including T cell receptor signaling, antigen recognition responses, and robust cytokine secretion capabilities. Jurkat cells are widely employed to dissect signal transduction pathways governing T cell activation, proliferation, and effector functions, making them a suitable host for studying genes involved in immune regulation and inflammatory signaling.
C15orf40, also designated as ILRUN, functions as a negative regulator of the canonical NF-??B pathway. Mechanistically, C15orf40 binds ubiquitin chains and associates with the autophagy receptors SQSTM1/p62 and NBR1 to inhibit phosphorylation of IKK?? (IKBKB), thereby stabilizing the inhibitor protein I??B?? (NFKBIA). This stabilization prevents NF-??B (RELA) nuclear translocation and suppresses transcription of pro-inflammatory cytokines. The pathway is activated by inflammatory stimuli such as TNF-??, IL-1??, and LPS, which converge on IKK?? activation. Disruption of C15orf40 relieves this inhibition, leading to enhanced NF-??B signaling and elevated expression of downstream targets including RELA and NFKBIA.
In Jurkat T lymphocytes, NF-??B is central to antigen-induced activation and cytokine production. Loss of C15orf40 in these cells is anticipated to potentiate NF-??B responses, providing a cellular platform to study the dynamics of negative feedback control in T cell-mediated immunity. This model is particularly relevant for exploring the molecular basis of inflammatory disorders and autoimmune diseases where unchecked NF-??B activity drives pathogenesis. The interaction of C15orf40 with p62 and NBR1 also links NF-??B regulation to autophagy pathways, enabling cross-talk studies between immune signaling and proteostasis.
Researchers can employ this knockout model in a variety of experimental settings, including NF-??B luciferase reporter assays to quantify pathway activity, western blotting for phosphorylated IKK?? and I??B?? to assess signaling kinetics, and RT-qPCR to measure pro-inflammatory cytokine transcript levels. Co-immunoprecipitation experiments can validate C15orf40 interactions with p62 and ubiquitin in T cells, while flow cytometry can monitor surface expression of activation markers such as CD25 and CD69. These cells are also suitable for genetic screens and drug testing aimed at identifying NF-??B inhibitors. For detailed validation data and technical support, please contact Ascent Research.
