ARFIP1 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Jurkat human T lymphocyte line, with targeted disruption of the ARFIP1 gene. This loss-of-function model enables dissection of ARFIP1 roles in membrane trafficking and actin remodeling. The polyclonal pool contains a heterogeneous mix of edited alleles, providing a robust population-level knockout effect without single-cell clone isolation. It is ideal for pooled functional assays, minimizing clonal artifacts.
The Jurkat cell line originates from peripheral blood of a 14-year-old male with acute lymphoblastic leukemia and serves as a model for TCR signaling, apoptosis, and T cell activation. Its well-characterized pathways and ease of culture make it a standard platform for lymphocyte biology and leukemia research. ARFIP1 knockout in this background allows assessment of how membrane trafficking impacts immune cell function.
ARFIP1 acts as an effector of the small GTPase ARF1, activated by GEFs such as GBF1. It binds to ARF1-GTP and PI4P, facilitating membrane tubulation and COPI vesicle coat recruitment at the Golgi. ARFIP1 interacts with ARF1, ARF3, ARF5, and possibly ARF6 and Rac1, linking ARF activation to actin cytoskeleton remodeling and Golgi maintenance. Key pathway constituents include PI4KIII??, the COPI complex, and actin.
In Jurkat T cells, ARFIP1 likely regulates membrane microdomains and receptor trafficking crucial for immune synapse formation. Knockout may alter TCR clustering and downstream signaling via ZAP70 and ERK. Given the leukemic origin, ARFIP1 disruption may also affect cancer cell proliferation and migration, offering insights into leukemogenesis and Golgi-dependent signaling.
Applications include western blotting and RT-qPCR for knockout confirmation, immunofluorescence for Golgi morphology, flow cytometry for CD69 and CD25, migration assays, co-immunoprecipitation for ARF1, and phospho-signaling analysis. This polyclonal knockout pool supports diverse studies in immune cell biology and membrane trafficking. For further details, contact Ascent Research.
