The EHD3 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated by disrupting the EHD3 gene in the Raji host cell line. This loss-of-function model serves as a versatile tool for investigating the endocytic recycling regulator EHD3 and its impact on receptor trafficking, cell adhesion, and signal transduction in a B lymphocyte context.
Raji is a human B lymphocyte cell line derived from a Burkitt lymphoma, characterized by Epstein-Barr virus (EBV) positivity and a lymphoblastoid phenotype. These suspension-adapted cells express B-cell surface markers and are widely employed in immunological studies, cancer research, and as a model for B-cell signaling pathways, offering a relevant system for studying hematopoietic malignancies and viral oncogenesis.
EHD3 encodes an EH domain-containing protein that orchestrates endocytic recycling by controlling the return of internalized receptors and adhesion molecules to the plasma membrane. It interacts with key endosomal regulators, including Rab4a, Rab11, Rabenosyn-5, and Arf6, and modulates the trafficking of integrins and growth factor receptors. Through these interactions, EHD3 influences downstream signaling pathways such as EGFR and Notch, and impacts cellular processes mediated by Rac1 and cell adhesion molecules, thereby regulating cell adhesion, migration, and proliferation.
In the Raji B-cell lymphoma model, disruption of EHD3 is anticipated to perturb integrin-dependent adhesion and migration, processes that are critical for lymphoma cell dissemination and potential drug resistance. Additionally, given the role of EHD3 in receptor recycling, this knockout model facilitates investigation of how endocytic trafficking regulates B-cell receptor signaling, antigen presentation, and interactions with the tumor microenvironment. The EBV-positive background further enables exploration of viral manipulation of host trafficking pathways.
Researchers can utilize this polyclonal knockout cell population in a range of assays to study endocytic recycling, cell migration, and signaling. Representative techniques include Western blotting and RT-qPCR for assessing EHD3 disruption, immunofluorescence to examine receptor localization, flow cytometry for measuring surface receptor recycling kinetics, and Transwell or wound healing assays to evaluate cell migration. The model is also suitable for phospho-signaling analysis and co-immunoprecipitation to map protein interactions. For additional information, please contact Ascent Research.
