The CD2AP Knockout Raji Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal cell population derived from the human Raji B lymphoblast line, featuring targeted disruption of the CD2AP gene. This loss-of-function model provides a versatile tool for studying CD2AP-dependent processes without the need for single-cell cloning, offering a heterogeneous pool of edited cells suitable for bulk functional assays and screening applications.
Raji cells are an Epstein-Barr virus (EBV)-positive B lymphocyte line originally isolated from a Burkitt’s lymphoma patient. They serve as a classical model for investigating B cell receptor (BCR) signaling, lymphomagenesis, and host-virus interactions. The cell line expresses surface immunoglobulins and key signaling components, making it an ideal host for examining immune synapse formation, antigen presentation, and cytoskeletal dynamics.
CD2AP encodes a multifunctional scaffold protein that couples membrane receptors to the actin cytoskeleton and endocytic machinery. It directly interacts with cortactin, CBL ubiquitin ligases, RAB4A, NPHS1 (nephrin), and PIK3R1 (p85) to orchestrate actin polymerization via the cortactin-ARP2/3 pathway, receptor internalization through Rab GTPase recruitment, and signal attenuation by promoting CBL-mediated ubiquitination. Upstream, CD2AP is activated by nephrin engagement, epidermal growth factor receptor (EGFR) signaling, and c-Cbl phosphorylation, positioning it as a hub for integrating cytoskeletal remodeling with vesicular trafficking.
In the Raji B cell context, CD2AP knockout disrupts BCR-mediated signaling and endocytosis, impairing immune synapse formation and antigen presentation. Loss of CD2AP alters actin cytoskeleton dynamics and may affect cell adhesion and migration. Given the EBV-positive background, this model also enables exploration of viral manipulation of host trafficking pathways, as CD2AP intersects with ubiquitination and endosomal sorting processes that are often co-opted by EBV oncoproteins.
Typical research applications include Western blotting for knockout validation, flow cytometry to assess surface receptor levels, and immunofluorescence to visualize actin reorganization. Co-immunoprecipitation and phospho-signaling arrays can map altered protein networks and BCR activation kinetics. Endocytosis assays using fluorescent ligands allow functional readouts. This polyclonal pool is particularly useful for pooled CRISPR screens, drug target validation, and dissection of signaling nodes linked to lymphoma biology and Alzheimer??s disease. For additional information or custom engineering services, please contact Ascent Research.
