The EHD4 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the EHD4 gene has been disrupted. This polyclonal pool contains a heterogeneous mixture of edited alleles, providing a robust loss-of-function model free from clonal selection bias. The knockout enables investigation of gene function in a population context, capturing the diversity of CRISPR-induced modifications.
Raji is a human B lymphocyte line derived from a Burkitt??s lymphoma patient, characterized by mature B cell markers such as surface immunoglobulin, CD19, and complement receptors. Rapidly proliferating and easy to culture, Raji cells serve as a widely used model for B cell biology, antibody production, and lymphomagenesis, retaining key signaling pathways downstream of the B cell receptor and receptor tyrosine kinases.
EHD4 (EH domain-containing protein 4) is an ATPase that orchestrates endocytic recycling by governing the transport of internalized receptors from early and recycling endosomes back to the plasma membrane. It functions downstream of receptor tyrosine kinases, such as EGFR, and in concert with Rab11, Arf6, and the EHD1 paralog. Through interactions with clathrin and the AP-2 complex, EHD4 coordinates cargo selection and vesicle formation. By controlling the surface availability of signaling receptors, EHD4 fine-tunes signal transduction cascades; for instance, sustained ERK phosphorylation depends on EHD4-mediated EGFR recycling. Consequently, EHD4 knockout disrupts recycling, dampening proliferative and survival signals.
Within Raji B lymphocytes, EHD4 is critical for the dynamic re?distribution of immune receptors, including the B cell receptor and cytokine receptors, which are essential for normal antibody responses and can be subverted in lymphomas. Eliminating EHD4 in this polyclonal knockout model allows systematic analysis of how endocytic recycling defects perturb BCR signaling, antigen uptake, and downstream transcriptional programs. The oncogenic context of Raji cells makes this system particularly relevant for investigating the role of membrane trafficking in Burkitt??s lymphoma pathogenesis and for identifying vulnerabilities in endocytic pathways that may be exploited therapeutically. Moreover, the model can be extended to study autoimmune conditions where dysregulated receptor recycling contributes to B cell hyperactivity.
Researchers can employ this knockout product in a range of assays. Western blotting and immunofluorescence confirm EHD4 loss and monitor recycling endosome markers. Flow cytometry quantifies surface levels of receptors such as EGFR or transferrin receptor, providing a direct measure of recycling efficiency. Functional analyses, including transferrin recycling kinetics and phospho-ERK time-course experiments, delineate the impact on trafficking and signaling. The cells are also suitable for co?culture with T cells or stromal cells to examine immune synapses, and for xenograft models to evaluate tumorigenicity. For technical inquiries and custom requests, contact Ascent Research.
