The ANXA1 Knockout HEK293T Polyclonal Cells are a polyclonal population of HEK293T human embryonic kidney epithelial cells in which CRISPR/Cas9-mediated gene disruption has been introduced at the ANXA1 locus. This polyclonal knockout model provides a heterogeneous loss-of-function system for investigating the multifunctional roles of annexin A1, avoiding the artifacts of clonal selection.
HEK293T cells are derived from HEK293 cells and stably express the SV40 large T antigen, enabling episomal replication of plasmids containing the SV40 origin and yielding high-titer viral packaging and robust protein expression. Their epithelial morphology, high transfectability, and widespread use in cell biology and signal transduction make them an ideal host for this knockout model.
ANXA1 encodes annexin A1, a calcium-dependent phospholipid-binding protein that mediates anti-inflammatory effects by inhibiting cytosolic phospholipase A2 (cPLA2) activity and decreasing prostaglandin E2 (PGE2) synthesis. It suppresses NF-??B and MAPK/ERK signaling through formyl peptide receptor 2 (FPR2/ALX), interacting with S100A11, EGFR, and NF-??B p65. Upstream, ANXA1 is transcriptionally activated by the glucocorticoid receptor (NR3C1) and regulated by cytokines including IL-6, TNF-??, and TGF-??. Downstream, ANXA1 attenuates ERK1/2 phosphorylation, I??B?? degradation, and NF-??B p65 activity, while also interfering with EGFR transactivation and binding to actin to regulate phagocytosis.
In the HEK293T epithelial context, ANXA1 knockout allows detailed examination of its functions in cell proliferation, apoptosis, and migration, with direct implications for cancer research focusing on breast and pancreatic malignancies. The model enables interrogation of glucocorticoid receptor-driven anti-inflammatory pathways and NF-??B signaling kinetics, providing insights into inflammatory and autoimmune disorders and cross-talk between arachidonic acid metabolism and growth factor signaling.
Key applications include Western blotting, RT-qPCR, and ELISA for confirming ANXA1 ablation and analyzing downstream targets; NF-??B luciferase reporter assays; phospholipase A2 activity assays; and Transwell migration assays to study invasive potential. Immunofluorescence and flow cytometry for surface FPR2 permit investigation of receptor internalization and signaling. This polyclonal knockout population is thus suited for immunomodulation studies, cancer invasion research, and dissection of glucocorticoid signaling cascades. For additional information or technical assistance, please contact Ascent Research.
