The EHBP1 Knockout Jurkat Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the EHBP1 gene has been disrupted in the human Jurkat T-lymphocyte line. This gene-disrupted pool provides a versatile loss-of-function model for investigating EHBP1-dependent endosomal trafficking, actin dynamics, and their integration with signal transduction. As a polyclonal knockout population, the product captures a spectrum of editing events, offering a representative system for functional genomics without clonal selection artifacts.
Jurkat cells are an immortalized CD4+ T-lymphoblast line derived from acute T-cell leukemia. They are widely used to study T-cell receptor (TCR) signaling, immune synapse formation, and adaptive immunity. The Jurkat background expresses key components of endocytic and actin regulatory pathways, making it a suitable host for dissecting EHBP1 functions in a lymphocyte context. These cells retain responsiveness to stimuli such as TCR crosslinking and are amenable to genetic perturbation and biochemical assays.
EHBP1 encodes an adaptor protein that bridges EH domain-containing proteins EHD1/EHD2 and F-BAR domain proteins syndapin I (PACSIN1) and syndapin II (PACSIN2), thereby coupling endocytic vesicle fission and recycling to N-WASP-mediated actin polymerization. This scaffold facilitates cargo sorting, receptor recycling, and directional cell migration. EHBP1 operates downstream of receptors including the insulin receptor and TCR, controlling processes such as GLUT4 translocation and actin reorganization. In T cells, EHBP1 may link TCR engagement to endosomal trafficking and cytoskeletal remodeling required for immune synapse function.
In Jurkat cells, disruption of EHBP1 allows interrogation of how endocytic machinery interfaces with actin dynamics during T-cell activation and migration. The knockout polyclonal population can be used to dissect the contribution of EHBP1 to transferrin uptake, chemokine-driven motility, and receptor surface expression. Altered phospho-Akt signaling downstream of TCR or insulin stimuli can be monitored, revealing EHBP1??s role in coordinating trafficking with signaling outputs. This model thus enables systematic study of EHBP1 loss on lymphocyte behavior.
Key applications include confocal imaging of actin and endosomal markers, flow cytometric analysis of receptor recycling, transferrin internalization assays, and immunoprecipitation with anti-EHD antibodies to probe complex assembly. The knockout cells are suited for migration assays and for investigating crosstalk between endocytosis and insulin signaling via phospho-Akt readouts. These tools support research into type 2 diabetes, cancer cell invasion, and immunodeficiency where EHBP1-associated trafficking is disrupted. For additional technical information, please contact Ascent Research.
