The ACAP2 Knouckout Jurkat Polyclonal Cells are a heterogeneous population of Jurkat T lymphocytes with CRISPR/Cas9-mediated disruption of the ACAP2 gene. This polyclonal knockout pool avoids clonal artifacts and provides a robust loss-of-function model for studying ACAP2-dependent membrane trafficking and signaling. The cells are derived from the Jurkat host line and are supplied as a ready-to-use population, suitable for immediate functional assays requiring ACAP2 deficiency.
The Jurkat cell line is an immortalized human T lymphocyte line established from the peripheral blood of a 14-year-old male with acute lymphoblastic leukemia. As suspension-growing lymphoblasts, Jurkat cells are widely used to investigate T cell receptor signaling, apoptosis, and immune cell biology. Their leukemic origin and well-characterized signaling networks make them ideal for genetic perturbation studies, including CRISPR-mediated gene knockout, to probe pathways governing T cell activation, adhesion, and migration.
ACAP2 is an ARF6 GTPase-activating protein that inactivates ARF6-GTP, thereby promoting clathrin-mediated endocytosis and integrin ??1 recycling. It is recruited to PIP3-rich membranes downstream of EGFR and Akt/PKB signaling. As a scaffold, ACAP2 interacts with clathrin heavy chain, the AP-2 complex, cortactin, and GULP1, linking endocytic machinery to actin dynamics. This suppresses ARF6-driven cytoskeletal remodeling and downstream Rac1 activity, coupling receptor signaling to cell adhesion and migration.
In Jurkat cells, ACAP2 disruption is predicted to alter integrin recycling and actin reorganization, impacting T cell adhesion, migration, and immune synapse formation. Since ACAP2 is implicated in cancer cell invasion and metastasis, this knockout model enables the study of ARF6-dependent trafficking in a T-ALL context, offering insights into leukemic cell dissemination and potential therapeutic targets. The model also facilitates examination of cross-talk between EGFR/Akt pathways and ARF6-mediated cytoskeletal changes.
Researchers can use these cells in flow cytometry to quantify surface integrin ??1, western blotting to monitor ARF6 and Akt phosphorylation, and immunofluorescence microscopy to visualize clathrin puncta and F-actin. Functional assays such as transwell migration, transferrin uptake, and cell adhesion assays provide quantitative measures of endocytosis and motility, while co-immunoprecipitation and GTPase activity assays assess protein interactions and ARF6 activation. These applications support studies in T cell biology, cancer metastasis, and ARF6 signaling. For further information, please contact Ascent Research.
