The GIT2 Knockout Raji Polyclonal Cells are a human B-cell loss-of-function model generated by CRISPR/Cas9-mediated disruption of the GIT2 gene in Raji lymphoblastoid cells. This polyclonal knockout cell population enables studies of GIT2-dependent processes without the confounding influence of clonal selection artifacts. The knockout product is provided as a ready-to-use polyclonal pool, facilitating robust and reproducible experimental designs in suspension-based B-cell systems.
Raji cells are an EBV-positive continuous B lymphoblastoid line derived from a Burkitt??s lymphoma patient. They grow in suspension, express surface immunoglobulin M (IgM) and B-cell receptors (BCR), and retain key features of antigen-presenting cells, including expression of ICAM-1 and integrin LFA-1. This cell line is widely employed as a model for humoral immunity, receptor signaling, and lymphoma cell biology, making it an ideal host for interrogating the role of adaptor proteins that coordinate adhesion and signaling.
GIT2 (G protein-coupled receptor kinase-interacting ArfGAP 2) is a multifunctional scaffold and GTPase-activating protein that localizes to focal adhesions and endocytic sites. It directly interacts with paxillin and the guanine nucleotide exchange factor ??-PIX, forming a structural platform that couples Arf6-mediated membrane trafficking to PAK/PIX-dependent cytoskeletal reorganization. GIT2 is regulated by B-cell receptor engagement, chemokine receptors such as CXCR4, integrin ligands, and GRK-mediated phosphorylation; it modulates the activity of Arf1, Arf6, PAK1, and Rac1, thereby controlling actin dynamics, adhesion complex turnover, and receptor recycling. Through its GIT-PIX-paxillin complex, GIT2 integrates signals from adhesion molecules to orchestrate lymphocyte polarity and migration.
In the Raji B-cell context, disruption of GIT2 perturbs BCR-stimulated signaling, impairs integrin inside-out activation (e.g., LFA-1 clustering), compromises firm adhesion to ICAM-1 or fibronectin, and reduces chemotactic migration toward SDF-1. This model thereby facilitates dissection of the molecular links between antigen receptor stimulation, GTPase regulation, and cytoskeletal remodeling, and provides a platform to investigate GIT2 contributions to lymphoma cell dissemination and immune synapse formation.
The knockout cells are well-suited for Western blotting and co-immunoprecipitation of GIT2 with ??-PIX and paxillin. Functional studies include static adhesion to ICAM-1 or fibronectin, transwell migration toward SDF-1, and immunofluorescence for F-actin and paxillin. Flow cytometry can quantify LFA-1 activation after BCR crosslinking, while phospho-protein analysis probes PAK and Rac1 signaling. The model also supports drug sensitivity testing for anti-lymphoma compounds. For additional information, please contact Ascent Research.
