The EHD2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte line, engineered to disrupt the EHD2 gene. This stable loss-of-function model overcomes limitations of transient knockdown approaches, providing a reliable system for investigating EHD2-dependent endocytic recycling and caveolae stabilization. The polyclonal format ensures genetic diversity while maintaining consistent EHD2 ablation, suitable for functional studies where clonal variability is undesirable.
The parental Raji cell line originates from Burkitt??s lymphoma and is Epstein?CBarr virus immortalized, retaining key B cell functions such as antibody production, antigen presentation, and immune surveillance. These lymphoblastoid cells are well-characterized models for studying B cell receptor (BCR) signaling and membrane trafficking, with endogenous expression of caveolae and actin machinery, making them an appropriate host for dissecting EHD2-mediated pathways.
EHD2 oligomerizes at caveolae necks, coupling membrane curvature to the actin cytoskeleton via interactions with caveolin-1 and cavin-1. Its activity is regulated by phosphatidylinositol 4,5-bisphosphate and membrane curvature, and it further assembles with PACSIN2, amphiphysin-1, EHBP1, actin, and myosin-1c to coordinate membrane tubulation and receptor recycling. Disruption of EHD2 impairs caveolae stabilization and clathrin-independent endocytosis, affecting downstream integrin recycling and cytoskeletal reorganization, which are linked to metabolic syndrome and myopathy.
In Raji B lymphocytes, EHD2 knockout provides a valuable tool to probe caveolae-dependent BCR internalization and antigen trafficking. Loss of EHD2 likely destabilizes caveolae, leading to altered receptor turnover and integrin dynamics that may impact B cell adhesion, migration, and immune function. Given the lymphomal background, this model also enables exploration of caveolae in cancer cell membrane dynamics and signaling.
Typical applications include endocytosis assays, immunofluorescence for caveolae and actin, co-immunoprecipitation of caveolar complexes, and western blotting. Flow cytometry can assess surface receptor levels, while migration assays evaluate cytoskeletal function. These cells support investigations into EHD2 biology in B cell receptor trafficking, metabolic disease mechanisms, and caveolae-targeted therapeutic development. For further details, contact Ascent Research.
