NECAP2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Burkitt lymphoma B lymphocyte cell line Raji, featuring targeted disruption of the NECAP2 gene. This loss-of-function model is designed to investigate the regulatory roles of NECAP2 in clathrin-mediated endocytosis and receptor trafficking within a B-cell context. The polyclonal format provides a heterogeneous knockout population, enabling studies that reflect a range of gene-disruption efficiencies without the bias of single-cell cloning.
The Raji parental cell line is an Epstein-Barr virus (EBV)-positive B lymphocyte model extensively used for studying B-cell malignancies and immune signaling. Raji cells exhibit constitutive NF-??B activation and grow in suspension, making them suitable for large-scale endocytosis and signal transduction assays. Their derivation from Burkitt lymphoma positions them as a relevant system for exploring oncogenic pathways and antigen presentation mechanisms.
NECAP2 encodes a phosphoinositide-binding endocytic protein that functions as a negative regulator of clathrin-mediated endocytosis by sequestering the AP-2 adaptor complex. Molecularly, NECAP2 interacts with AP2A2, AP2B1, clathrin heavy chain (CLTC), and phosphoinositides, influencing coat assembly and vesicle formation. Upstream, its activity is modulated by phosphoinositide signaling and B cell receptor (BCR) activation. Downstream, NECAP2 loss-of-function is predicted to enhance clathrin-coated vesicle formation, thereby altering cell surface receptor expression, AP-2 complex localization, and internalization of endocytic cargo such as receptor tyrosine kinases.
In Raji B lymphocytes, NECAP2 knockout is expected to perturb BCR trafficking and downstream signaling cascades, including those mediated by NF-??B, thereby impacting immune functions like antigen presentation and antibody production. This model allows dissection of how endocytic dysregulation contributes to lymphoid malignancy phenotypes. Researchers can leverage the constitutive NF-??B background to assess cross-talk between endocytosis and survival pathways.
Typical applications include mechanistic studies of endocytosis and receptor trafficking using transferrin uptake assays, immunofluorescence for AP-2 localization, and co-immunoprecipitation of endocytic complexes. The cells are also suited for functional genomics screens, flow cytometric analysis of surface receptor levels, RNA-seq transcriptomics following BCR stimulation, and drug sensitivity profiling in lymphoma. For additional details, please contact Ascent Research.
