The ADAM17 Knockout Jurkat Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal population derived from the Jurkat T lymphocyte line, designed for targeted disruption of the ADAM17 gene. This loss-of-function model provides a heterogeneous knockout background for investigating ADAM17-mediated ectodomain shedding without clonal selection, enabling studies of gene dosage effects and pathway attenuation in a T cell context.
Jurkat cells are an immortalized human T lymphocyte model originally established from a patient with acute T cell leukemia. Widely used to explore T cell receptor signaling, apoptosis, and HIV infection, Jurkat cells offer a robust platform for genetic manipulation due to their well-defined signaling networks and rapid proliferation.
ADAM17 is a zinc-dependent metalloprotease that functions as a sheddase for numerous transmembrane substrates, including pro-TNF-alpha, EGFR ligands (TGF-alpha, HB-EGF, amphiregulin, epiregulin), L-selectin, IL-6R, APP, and Notch1. Its activity is regulated by upstream factors such as PMA, TNF-alpha, IL-1beta, EGF, and lipopolysaccharide, often requiring adaptor proteins iRhom1 or iRhom2 for maturation and trafficking. Substrate cleavage releases soluble ectodomains that activate downstream pathways: TNF-alpha engages TNF-R1 to stimulate NF-kB; EGFR ligands trigger MAPK/ERK signaling; IL-6R shedding permits IL-6 trans-signaling via JAK/STAT. The tissue inhibitor TIMP-3 provides endogenous negative regulation. Consequently, ADAM17 orchestrates a wide range of cellular responses, coupling environmental cues to inflammatory cytokine production, growth factor signaling, and cell fate decisions.
In Jurkat T cells, ADAM17 knockout disrupts the constitutive and inducible shedding of these ligands, providing a clean background to study TNF-alpha and EGFR ligand processing in an immune context. The polyclonal knockout population enables analysis of how impaired ectodomain release alters NF-kB and MAPK pathway activation, and downstream transcriptional events. Researchers can quantify changes in surface receptor levels, cytokine secretion, and cell viability, linking ADAM17 function to T cell activation, apoptosis, and leukemia biology. Moreover, this model permits the investigation of compensatory shedding mechanisms and gene dosage effects.
This product is suited for mechanistic studies of TNF-alpha-driven inflammation, EGFR transactivation in immune cells, and the evaluation of ADAM17 as a therapeutic target in rheumatoid arthritis, inflammatory bowel disease, psoriasis, and cancer. Common readouts include Western blotting for cleaved substrates, ELISA for shed TNF-alpha, flow cytometry for receptor surface expression, phospho-specific flow cytometry for signaling, RT-qPCR for transcriptional targets, co-immunoprecipitation for interactors, and apoptosis assays. For additional technical information and protocols, please contact Ascent Research.
