The AFAP1 Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to ablate AFAP1 gene expression in the Jurkat T lymphocyte line. This heterogeneous population, generated via CRISPR/Cas9-mediated gene disruption without single-cell cloning, provides a physiologically relevant loss-of-function model for studying AFAP1-dependent mechanisms. Its polyclonal nature ensures representation of diverse genetic backgrounds and minimizes clonal artifacts.
Jurkat cells are a well-characterized immortalized human T lymphocyte line derived from the peripheral blood of a 14-year-old male with acute lymphoblastic leukemia. They express T cell receptors and the IL-2 receptor, serving as a canonical model for T cell signaling, activation, and immune synapse formation. Their genetic tractability enables robust CRISPR/Cas9 genome editing, making them ideal for dissecting molecular pathways underlying T cell function and leukemogenesis.
AFAP1 encodes an adaptor protein that bridges Src family kinases with the actin cytoskeleton. It interacts with cortactin, vinculin, Src, and PKC??, and is activated downstream of integrin receptors and receptor tyrosine kinases. Upon integrin engagement, AFAP1 recruits Src kinases to focal adhesions, promoting phosphorylation of focal adhesion kinase (FAK) and downstream activation of RhoA/ROCK and ERK signaling. This cascade drives actin polymerization, focal adhesion maturation, and cell migration. AFAP1 also serves as a scaffold for PKC??-mediated signaling, further modulating cytoskeletal dynamics.
In the Jurkat T cell host, AFAP1 is critical for integrin ??5??1?Cmediated adhesion and immune synapse formation. Knockout of AFAP1 may impair Src-dependent signaling at focal adhesions, disrupt RhoA-dependent actin remodeling, and alter T cell spreading and migration. These defects can compromise antigen recognition and T cell activation, providing a platform to dissect the molecular basis of immune dysregulation. The polyclonal knockout population allows assessment of heterogeneous responses that more closely mimic physiological variability.
Typical applications include western blotting for phospho-Src and actin reorganization, immunofluorescence localization of vinculin and focal adhesions, transwell migration assays, and flow cytometry for integrin expression. Co-immunoprecipitation studies can verify disrupted AFAP1?CSrc complexes. This model is invaluable for research in T cell biology, cancer metastasis, and acute lymphoblastic leukemia. For further details, contact Ascent Research.
