The DNAJC1 Knockout Raji Polyclonal Cells comprise a heterogeneous population of Raji B lymphoblastoid cells in which the DNAJC1 gene has been disrupted using CRISPR/Cas9 technology. This polyclonal knockout model provides a loss-of-function tool to investigate the cellular consequences of DNAJC1 deficiency in a B-cell lymphoma background. As a polyclonal pool, it captures a range of editing events across the cell population, enabling robust functional studies without clonal selection artifacts.
The parental Raji cell line is an EBV-immortalized B lymphocyte line derived from a Burkitt lymphoma patient. These cells exhibit a mature B-cell phenotype with high secretory pathway activity, rendering them inherently sensitive to perturbations in endoplasmic reticulum (ER) homeostasis. Their lymphoblastoid nature and well-characterized response to ER stressors make them an ideal host for dissecting ER proteostasis mechanisms relevant to B-cell malignancies.
DNAJC1 encodes an ER-resident Hsp40 co-chaperone that directly interacts with HSPA5 (BiP) and stimulates its Hsp70 ATPase activity, thereby facilitating protein folding and ER-associated degradation (ERAD). Upstream ER stress signals transmitted via ATF6, ERN1 (IRE1), and EIF2AK3 (PERK) engage this chaperone network. In the knockout cells, impaired BiP co-chaperone function disrupts ER quality control, leading to accumulation of misfolded proteins and activation of the unfolded protein response (UPR). Downstream, this triggers transcriptional reprogramming exemplified by DDIT3 (CHOP) upregulation and XBP1 splicing, shifting the cellular balance toward apoptosis under unresolved stress.
In the lymphoma context, DNAJC1 disruption is particularly significant because Raji cells rely on robust ER capacity to sustain rapid proliferation and immunoglobulin synthesis. Loss of DNAJC1 sensitizes these cells to ER stress-induced apoptosis, potentially uncovering co-chaperone dependencies in B-cell lymphomagenesis. This model thus provides a platform to study how ER stress modulators influence survival decisions in Burkitt lymphoma and other B-cell malignancies.
Typical applications include quantitative profiling of UPR markers by Western blotting (HSPA5, CHOP, XBP1s) and RT-qPCR, functional assays for apoptosis (flow cytometry with ER stress inducers like tunicamycin or thapsigargin), and co-immunoprecipitation studies with HSPA5 or SIL1 to probe co-chaperone interaction networks. High-content screens for ER stress modulators and transcriptomic analyses by RNA-seq are also facilitated. For further technical details or to discuss experimental design, please inquire with Ascent Research.
