The AAGAB Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human AAGAB gene. Produced by CRISPR/Cas9-mediated gene disruption in Jurkat T lymphocytes, this loss-of-function model yields a heterogeneous pool of cells with targeted disruptions that impair AAGAB expression, suitable for population-based functional studies without clonal selection bias.
Jurkat cells, derived from the peripheral blood of a 14-year-old male with acute T cell leukemia, are a widely used human T lymphocyte model for studying TCR signaling, apoptosis, and immune responses. Their high transfectability and rapid growth make them ideal for genetic manipulation and downstream biochemical assays in immunological research.
AAGAB encodes an alpha- and gamma-adaptin binding protein that facilitates clathrin-mediated endocytosis (CME) by interacting with the AP-2 adaptor complex. It directly binds AP2A1, AP2A2, and AP1G1, and associates with clathrin heavy chain (CLTC) to drive vesicle coat assembly. Upstream regulators include cellular stress signals, phosphoinositides, and kinases such as GAK, while downstream events involve AP-2 complex recruitment and internalization of receptors like transferrin and EGFR. The mechanistically linked components AP2A1, AP2A2, AP1G1, CLTC, DNM2, FCHO1/2, and Eps15 underscore its integrated role in endocytic trafficking.
In Jurkat T cells, AAGAB knockout disrupts CME, impairing AP-2 function and reducing internalization of cell surface receptors. This perturbation can alter TCR endocytosis and signaling dynamics, impacting T cell activation, desensitization, and immune response modulation. The Jurkat background provides a physiologically relevant T lymphocyte system to examine how endocytic dysregulation affects adaptive immunity, with the polyclonal knockout format capturing a range of editing events that facilitate phenotype identification.
Representative assays include TCR downregulation to quantify receptor internalization kinetics, transferrin uptake to measure CME efficiency, and EGFR endocytosis for general endocytic capacity. Complementary techniques such as Western blotting, clathrin immunofluorescence, and co-immunoprecipitation enable detailed biochemical characterization, while RNA-seq reveals transcriptional network adaptations. These cells are valuable for modeling punctate palmoplantar keratoderma type I cellular defects and investigating endocytic dysregulation in cancer. For additional information or to request a custom knockout, contact Ascent Research.
