AFTPH Knockout Jurkat Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the AFTPH gene, which encodes aftiphilin, in the Jurkat human T lymphocyte line. This model enables investigation of AFTPH-dependent clathrin-mediated endocytosis and intracellular trafficking pathways. The polyclonal format provides a heterogeneous pool for robust population-level studies, avoiding clonal selection biases. It is intended for advanced biomedical research requiring reliable AFTPH loss-of-function in a T cell context.
The Jurkat line, derived from peripheral blood of a 14-year-old male with acute T cell leukemia, is an immortalized T lymphocyte model extensively utilized for studies of TCR signaling, apoptosis, and leukemia biology. Its robust suspension growth, fast doubling time, and comprehensive characterization of signaling cascades provide an optimal backdrop for probing the endocytic machinery underlying immune cell function and transformation.
Aftiphilin (AFTPH) serves as a critical clathrin-adaptor scaffold, physically linking AP-1 (??-adaptin) and AP-2 (??-adaptin) complexes with clathrin heavy chain to orchestrate coated vesicle formation at the trans-Golgi network and plasma membrane. Its regulation involves ARF GTPases, the phosphoinositide PIP2, and the kinase AAK1, and it drives internalization of diverse cargo including the transferrin receptor, EGFR, and the TCR/CD3 complex. Beyond endocytosis, AFTPH participates in endosomal sorting, lysosomal enzyme trafficking, and likely intersects with the ESCRT pathway, positioning it at a nexus of membrane transport processes.
In the Jurkat T cell context, AFTPH knockout is anticipated to block clathrin-dependent internalization of surface receptors, most prominently the TCR/CD3 complex, thereby suppressing downstream signal transduction (e.g., ZAP70 and ERK phosphorylation). The known interaction of AFTPH with HIV-1 Gag and ESCRT-I components suggests these knockout cells may also exhibit defective viral egress, making them a valuable tool for studying host factors in HIV-1 replication. Furthermore, the model permits exploration of how endocytic deficits contribute to T cell leukemia pathogenesis and lysosomal storage disorders.
This polyclonal knockout cell product is optimized for quantitative assays such as transferrin uptake kinetics and flow cytometric analysis of surface TCR/CD3 expression. It is equally valuable for biochemical studies, including co-immunoprecipitation of AFTPH with AP-1 and AP-2 complexes, western blotting for aftiphilin and clathrin adaptors, and confocal immunofluorescence colocalization with endosomal markers like LAMP1. Researchers can employ it for HIV-1 release assays to probe host factor dependencies, transcriptomic profiling via RNA-seq, and phospho-flow cytometry to assess TCR signaling intermediates. For technical specifications or purchasing information, please contact Ascent Research.
