The DNAJC10 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes harboring targeted disruption of the DNAJC10 gene. This loss-of-function model enables investigation of DNAJC10/ERdj5 function without relying on pharmacological inhibition or transient silencing approaches. The polyclonal format provides a heterogeneous knockout pool that reflects the genetic diversity inherent to CRISPR-mediated gene disruption, making it suitable for population-level functional studies where clonal homogeneity is not essential.
Raji cells are a well-characterized human B lymphocyte line derived from a Burkitt??s lymphoma patient and are Epstein?CBarr virus (EBV)-positive. These cells retain many features of mature B cells, including active immunoglobulin production and robust secretory pathway activity, rendering them particularly relevant for studying endoplasmic reticulum (ER) homeostasis and protein quality control. Their lymphomatous origin also makes Raji cells a valuable model for investigating the intersection of ER stress, UPR signaling, and B cell malignancies.
DNAJC10 encodes ERdj5, an ER-resident Hsp40 co-chaperone and disulfide reductase. ERdj5 reduces disulfide bonds in misfolded glycoproteins, cooperating with EDEM1, OS9, HSPA5/BiP, SEL1L, and SYVN1/HRD1 to facilitate substrate retrotranslocation and proteasomal degradation via ERAD. Its activity is induced by ER stress agents like tunicamycin and thapsigargin, downstream of UPR sensors ERN1/IRE1, ATF6, and EIF2AK3/PERK. Disruption of DNAJC10 leads to accumulation of reduced, misfolded proteins and impaired ERAD clearance.
In Raji B cells, DNAJC10 knockout severely compromises ERAD efficiency, resulting in constitutive ER stress and heightened basal UPR activity. This phenotype is particularly informative for dissecting the adaptive and pro-apoptotic branches of the UPR in a lymphoid context. Because elevated ER stress and UPR activation are hallmarks of aggressive lymphomas and contribute to chemoresistance, this knockout model can be employed to examine how ERAD deficiency influences B cell survival, proliferation, and drug sensitivity. Additionally, as professional secretory cells, Raji B lymphocytes depend heavily on protein quality control; thus, this model offers insights into how ERAD defects alter immunoglobulin folding and secretion.
The DNAJC10 Knockout Raji Polyclonal Cells are suitable for a wide range of experimental approaches. Western blotting and RT-qPCR can monitor UPR markers (e.g., HSPA5, spliced XBP1) and ERAD substrates. Flow cytometry enables assessment of viability, apoptosis (with/without tunicamycin or thapsigargin), and surface immunoglobulin. Co-immunoprecipitation and ubiquitination assays characterize interactions with BiP, EDEM1, or SYVN1 and substrate ubiquitination. Immunofluorescence visualizes ER morphology. Combined with MG132, this model dissects ERAD kinetics. These cells are a powerful tool for ER proteostasis, UPR signaling, protein misfolding, and B cell lymphomagenesis research. For further details, please contact Ascent Research.
