The ERP44 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte line, designed to disrupt the ERP44 gene. This polyclonal knockout model preserves population heterogeneity, allowing robust functional analyses without clonal selection. The heterogeneous pool of cells with targeted gene disruption enables loss-of-function studies in a lymphoma-relevant background, facilitating investigation of ERP44-dependent processes.
The Raji cell line is an EBV-positive lymphoblastoid line originally established from a male patient with Burkitt’s lymphoma. Widely used as a model for B cell lymphomas and immune cell function, Raji cells provide a well-characterized platform for investigating oncogenic mechanisms and ER stress responses, providing a physiologically relevant context for studying B cell malignancies.
ERP44 encodes an endoplasmic reticulum (ER) resident thioredoxin family protein central to oxidative protein folding and calcium homeostasis. It interacts with ERO1L and PDIA2 to modulate disulfide bond formation and binds ITPR1 to regulate ER calcium release. Under ER stress, upstream transcription factors ATF6, XBP1, and ATF4 upregulate ERP44, linking it to UPR pathways. ERP44 forms complexes with chaperones HSPA5/BiP and ERP57, and its activity intersects with PERK/EIF2AK3?CCHOP/DDIT3 signaling. Thus, ERP44 coordinates protein quality control, UPR, and calcium dynamics, with disruption predicted to impair ER proteostasis and sensitize cells to stress-induced apoptosis.
In Raji B lymphocytes, ERP44 disruption perturbs ER?Ccalcium communication and compromises oxidative folding, likely exacerbating basal ER stress and sensitizing cells to apoptosis under proteotoxic conditions. This model helps dissect UPR-driven survival mechanisms in Burkitt lymphoma and other ER stress-associated diseases. The absence of ERP44 may also alter ITPR1-mediated calcium release dynamics, feeding back onto UPR signaling and influencing cell fate decisions in lymphoma cells, potentially revealing dependencies on alternative chaperone networks or calcium handling pathways that represent therapeutic vulnerabilities.
These polyclonal knockout cells enable diverse experimental approaches: Western blotting for UPR markers (CHOP, GRP78), RT-qPCR profiling of ER stress genes, and Annexin V flow cytometry quantify apoptotic sensitivity. Calcium imaging tracks altered ER?Ccytosolic fluxes, while tunicamycin or thapsigargin viability assays assess stress resilience. Co-immunoprecipitation can map shifted protein interactions. Moreover, the polyclonal nature ensures representation of diverse editing events, facilitating population-level studies of gene function. Applications span cancer biology, drug screening, and calcium signaling. For further details, contact Ascent Research.
