The AZI2 Knockout Jurkat Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted disruption of the AZI2 gene in the Jurkat T lymphoblastoid cell line. This heterogeneous population captures diverse gene-editing events, providing a versatile loss-of-function model for studying AZI2-dependent signaling pathways.
The parental Jurkat cell line (E6.1) is a human T lymphoblastoid cell line derived from the peripheral blood of a 14-year-old male with acute T-cell leukemia. Jurkat cells are widely employed as a model for T-cell receptor signaling, apoptosis, and innate immune responses. Their suspension growth and well-characterized signaling machinery make them particularly suited for high-throughput screening and biochemical analyses of immune activation pathways.
AZI2 (5-azacytidine-induced protein 2) is an essential adaptor protein that coordinates innate immune signaling downstream of pattern recognition receptors such as Toll-like receptor 4 (TLR4) and nucleotide-binding oligomerization domain-containing protein 2 (NOD2). Mechanistically, AZI2 interacts with the kinases TBK1 and IKK??, as well as the scaffold protein TANK, to facilitate phosphorylation and activation of the transcription factors IRF3 and IRF7. Concurrently, AZI2 contributes to NF-??B pathway activation through associations with the adaptors TRAF6 and RIPK2, ultimately promoting the expression of pro-inflammatory cytokines including TNF, IL-6, and IFN-??. In addition, AZI2 has been implicated in the regulation of autophagy. Knockout of AZI2 thus disrupts these critical signaling cascades, impairing innate immune gene induction and potentially altering autophagic flux.
In the Jurkat T-cell background, AZI2 deficiency principally affects the responsiveness to innate immune stimuli such as lipopolysaccharide (LPS) and muramyl dipeptide (MDP), which signal through TLR4 and NOD2, respectively. Jurkat cells express components of these pathways, and AZI2 knockout is expected to attenuate TLR- and NOD2-driven NF-??B and IRF activation. This model enables dissection of AZI2??s role in T-cell intrinsic innate immunity, where it may intersect with T-cell receptor signaling and contribute to inflammatory responses in autoimmune and infectious disease contexts. Moreover, the disruption of autophagy regulation may provide insights into the interplay between immune signaling and cellular homeostasis in leukemic T cells.
The AZI2 Knockout Jurkat Polyclonal Cells are intended for advanced biomedical research applications including the study of innate immune signaling, NF-??B pathway dynamics, autophagy, and viral infection models. Typical experimental workflows involve stimulation with TLR4 or NOD2 agonists followed by quantification of signaling outputs. Representative assays include Western blot analysis of phospho-TBK1, phospho-p65, and AZI2 levels; RT-qPCR for cytokine transcription (e.g., TNF, IL-6, IFN-??); NF-??B luciferase reporter assays; ELISA for secreted cytokines; flow cytometric detection of intracellular phospho-NF-??B; immunofluorescence imaging of p65 nuclear translocation; and co-immunoprecipitation to assess TBK1?CAZI2 interactions. These polyclonal cells are also suitable for RNA-seq-based transcriptomic profiling to map AZI2-dependent gene networks. For further information, please contact Ascent Research.
