The DNAJB9 Knockout Jurkat Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal population derived from the Jurkat human T lymphoblastoid cell line, engineered for disruption of the DNAJB9 gene. This heterogeneous pool of knockout cells enables studies of DNAJB9 loss-of-function without single-cell clonal expansion, maintaining functional diversity suitable for downstream assays. The CRISPR-mediated gene disruption creates a versatile model for investigating the roles of this ER co-chaperone in T-cell biology.
Jurkat cells, originally isolated from an acute T-cell leukemia patient, are a cornerstone model for T-lymphocyte signaling, apoptosis, and adaptive immunity. They harbor PTEN deficiency, which enhances oncogenic signaling and stress sensitivity, and constitutively produce IL-2 while maintaining functional TCR complexes. Their suspension growth and lymphoblastoid phenotype facilitate robust genetic manipulation and high-throughput analyses of cell-mediated immune responses and cytokine production.
DNAJB9 functions as an ER-resident Hsp40 co-chaperone integral to the unfolded protein response (UPR) and ER-associated degradation (ERAD). It recognizes misfolded proteins and recruits BiP (HSPA5) to promote refolding or target substrates for proteasomal elimination through interactions with VCP, SEL1L, and HRD1. DNAJB9 is transcriptionally induced by ER stress via IRE1/XBP1, PERK/ATF4, and ATF6 arms, and operates downstream of these sensors to mitigate proteotoxic stress. Disruption of DNAJB9 impairs substrate handling, leading to accumulation of misfolded proteins and sensitization to ER stress-induced apoptosis.
In Jurkat T-cells, DNAJB9 knockout offers a system to dissect the role of ER proteostasis in T-cell acute lymphoblastic leukemia (T-ALL) pathogenesis. The heightened secretory load and redox stress in leukemic lymphocytes make the UPR and ERAD pathways critical for survival and drug resistance. Loss of DNAJB9 allows investigation of co-chaperone contributions to apoptotic thresholds, TCR signaling, and cytokine output under proteotoxic challenge, thereby illuminating mechanisms of therapy resistance and stress adaptation in T-cell malignancies and beyond.
These polyclonal knockout cells support diverse experimental approaches: RT-qPCR for XBP1 splicing, Western blotting of UPR markers (BiP, ATF4, CHOP), and flow cytometry for apoptosis assessment. Co-immunoprecipitation with BiP or VCP reveals altered chaperone complexes, while immunofluorescence visualizes ER morphology and protein aggregation. Cell viability assays under ER stressors (thapsigargin, tunicamycin) and drug treatments facilitate studies of drug resistance and proteotoxicity. For further details, contact Ascent Research.
