This CRISPR/Cas9-edited polyclonal population of DNAJB11 knockout Raji cells provides a reliable loss-of-function model for studying endoplasmic reticulum biology. The cells carry targeted disruptions of the DNAJB11 gene, generating a heterogeneous mixture of knockout alleles within the population. This polyclonal format avoids the artifacts and bottlenecks of clonal selection, making it ideal for bulk biochemical and functional assays where population-level responses are of primary interest.
Raji cells are a human B lymphocyte line derived from an EBV-positive Burkitt lymphoma, widely employed in immunology and cancer research. They proliferate in suspension and retain characteristics of mature B cells, including surface immunoglobulin expression and active secretory pathways. Their genetic background and robust growth make them a suitable host for studying the effects of gene disruption on antibody production, signal transduction, and oncogenic processes.
DNAJB11 encodes an ER luminal co-chaperone that directly interacts with HSPA5 (BiP) to facilitate protein folding and ER-associated degradation (ERAD). It functions downstream of the unfolded protein response (UPR) sensors ATF6 and IRE1/XBP1, and is linked to the PERK/EIF2AK3/ATF4/DDIT3 axis. DNAJB11 cooperates with HSP90B1, DERL1, and ERdj family members to recognize misfolded substrates and target them for retrotranslocation. Its loss disrupts BiP recruitment and ERAD efficiency, causing accumulation of unfolded proteins and sustained UPR activation, ultimately sensitizing cells to ER stress-induced apoptosis.
In Raji B cells, which rely on a high-flux secretory pathway for immunoglobulin production, DNAJB11 knockout creates a disease-relevant model for ER stress-driven pathology. This system is particularly informative for autosomal dominant tubulointerstitial kidney disease research, where DNAJB11 mutations disturb renal epithelial proteostasis. The polyclonal setting preserves native heterogeneity and allows investigation of UPR dynamics, ERAD competency, and crosstalk with B cell receptor signaling, revealing how lymphoma cells cope with proteotoxic insults.
Researchers can utilize this knockout population for western blot analysis of UPR markers (HSPA5, ATF4, DDIT3), RT-qPCR detection of XBP1 splicing and UPR target genes, and apoptosis measurements via annexin V staining after thapsigargin or tunicamycin treatment. Flow cytometry enables assessment of surface immunoglobulin or viability changes, while functional assays probe ERAD activity or calcium flux. This model also supports drug screens targeting proteostasis or combination therapies in lymphoma. For further details, please contact Ascent Research.
