The EDEM2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from Raji B lymphocytes, featuring disruption of the EDEM2 gene. This polyclonal product provides a heterogeneous loss-of-function model for studying EDEM2-dependent processes without clonal selection bias, enabling robust functional interrogation of ER-associated degradation (ERAD) in a human B cell context.
Raji is an EBV-positive Burkitt’s lymphoma B lymphocyte line that serves as a well-established model for B cell malignancies and humoral immunity. These cells express surface immunoglobulin and secrete antibodies, offering a relevant system to investigate protein secretion pathways and ER stress responses. Their rapid proliferation and well-characterized signaling networks facilitate genetic manipulation and diverse downstream assays.
EDEM2 encodes a mannose-trimming lectin that acts in the ERAD pathway, recognizing misfolded glycoproteins and trimming mannose residues to promote their retrotranslocation and proteasomal degradation. Its expression is upregulated by the unfolded protein response (UPR) transcription factors ATF6 and XBP1 in response to ER stress inducers such as thapsigargin and tunicamycin, which activate the PERK/eIF2?? pathway. EDEM2 functions within a multi-protein complex that includes the E3 ubiquitin ligase HRD1, the adaptor SEL1L, and recognition factors OS9, XTP3-B, Derlin-1, and VIMP. Well-characterized ERAD substrates like CD3-delta and the alpha-1-antitrypsin Z variant accumulate when EDEM2 is disrupted, triggering sustained UPR signaling through PERK and IRE1 kinases.
In Raji B lymphocytes, EDEM2 knockout disrupts ERAD, causing retention of misfolded glycoproteins and heightened ER stress. This perturbs the secretory function of these antibody-producing cells, potentially triggering UPR-mediated apoptosis or altered surface marker expression. The model is thus critical for dissecting how ER quality control influences B cell survival, differentiation, and lymphomagenesis, especially in EBV-driven oncogenesis where ER stress pathways are often dysregulated.
These polyclonal knockout cells support diverse applications such as proteomic identification of EDEM2 substrates, mechanistic studies of ERAD complex assembly, and functional genomics of glycoprotein quality control. Assays including western blotting for ER stress markers, RT-qPCR of UPR target genes, flow cytometric analysis of apoptosis and CD3-delta degradation, and co-immunoprecipitation of ERAD components are highly relevant. The model also facilitates drug discovery targeting ER stress-related diseases, including B cell lymphomas and neurodegenerative disorders. For further technical details, please contact Ascent Research.
