The TNF Knockout Jurkat E6.1 Cell Line is a CRISPR/Cas9-edited knockout cell line featuring targeted disruption of the TNF gene in the Jurkat E6.1 T-lymphocyte background. This model provides a defined loss-of-function system for investigating TNF-dependent signaling pathways without interference from residual endogenous TNF expression. By eliminating TNF production, researchers can dissect its contributions to autocrine and paracrine signaling mechanisms in a well-characterized human immune cell context.
Jurkat E6.1 is an immortalized human T-cell line derived from acute T-cell leukemia, maintaining a CD4+ phenotype and suspension growth characteristics. Widely adopted as a model for T-cell activation and cytokine secretion, these cells respond to stimuli such as T-cell receptor (TCR) engagement and co-stimulation, leading to robust activation of transcription factors including NF-??B, NFAT, and AP-1. The Jurkat E6.1 background thus provides a physiologically relevant platform for studying how TNF integrates into T-cell signaling networks and influences immune responses.
TNF encodes tumor necrosis factor, a potent pro-inflammatory cytokine that exerts pleiotropic effects by binding to its receptors TNFR1 and TNFR2. Upon TNFR1 ligation, adaptor proteins TRADD and TRAF2 are recruited, initiating downstream signaling cascades that lead to activation of the I??B kinase (IKK) complex and subsequent NF-??B nuclear translocation, as well as stimulation of JNK and p38 MAPK pathways. These events drive transcription of NF-??B target genes such as IL6 and IL8. Alternatively, under conditions inhibiting NF-??B survival signals, TNF receptor engagement promotes assembly of a death-inducing complex comprising FADD and caspase-8, which triggers extrinsic apoptotic signaling through caspase-3 and cleavage of substrates like BID. Additional components like RIPK1 and MLKL connect TNF signaling to necroptotic cell death. The interplay between these pathways determines cell fate, balancing inflammation, survival, and apoptosis.
In T lymphocytes, TNF functions as both an autocrine and paracrine regulator, influencing activation thresholds, cytokine production, and apoptotic sensitivity. By ablating TNF expression in Jurkat E6.1 cells, this knockout line enables precise dissection of TNF-dependent vs. -independent signaling events following TCR stimulation or other immune challenges. It serves as a critical tool for studying the role of TNF in autoimmune pathologies such as rheumatoid arthritis, inflammatory bowel disease, and psoriasis, as well as in septic shock and cancer-related inflammation. The model allows researchers to distinguish contributions of T-cell-derived TNF from those of other immune and tissue cell types.
Researchers can employ the TNF Knockout Jurkat E6.1 Cell Line in a variety of experimental settings, including Western blot analysis of phospho-NF-??B and phospho-JNK to assess MAPK and NF-??B pathway activity, ELISA or intracellular cytokine staining to quantify IL-6 or IL-8 secretion, and flow cytometric Annexin V assays to measure apoptosis induction. The line is also suitable for NF-??B luciferase reporter assays and RT-qPCR quantification of downstream transcriptional targets. Furthermore, it provides an isogenic background for co-immunoprecipitation studies of TNFR1 complex assembly and for screening potential TNF inhibitors. For additional information, please contact Ascent Research.
