The DNAJC3 Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat immortalized human T lymphocyte line, engineered to disrupt the DNAJC3 gene. DNAJC3 encodes P58IPK, a co-chaperone that functions as a critical inhibitor of the double-stranded RNA-dependent protein kinase PKR (EIF2AK2) and as a modulator of the unfolded protein response (UPR) during endoplasmic reticulum (ER) stress. This polyclonal knockout product provides a heterogeneous pool of cells carrying diverse loss-of-function mutations, enabling robust functional genomics studies without the selective pressure of single-cell cloning. The use of CRISPR/Cas9-mediated gene disruption ensures efficient target-gene ablation, allowing researchers to dissect DNAJC3-dependent signaling networks in a well-characterized T-cell model system.
Jurkat cells are derived from the peripheral blood of a 14-year-old male with acute T-cell leukemia and represent a widely adopted model for investigating T-cell receptor signaling, apoptosis, and leukemogenesis. These suspension-adapted lymphoblastoid cells exhibit constitutive activation of multiple signaling cascades, including NF-??B, MAPK, and PI3K/AKT pathways, making them particularly suitable for probing the interplay between oncogenic signaling and stress responses. The immortalized nature of Jurkat cells facilitates long-term culture and high-throughput screening applications, while their hematopoietic origin offers a physiologically relevant context for studying the role of DNAJC3 in immune cell function and hematological malignancies.
At the molecular level, DNAJC3/P58IPK operates at the nexus of the integrated stress response and UPR. Under basal conditions, P58IPK binds to PKR and prevents its autophosphorylation, thereby suppressing eIF2?? phosphorylation and maintaining cap-dependent translation initiation. Upon ER stress induction by agents such as tunicamycin or thapsigargin, P58IPK is transcriptionally upregulated by ATF6 and XBP1, and its protein product interacts with the ER-resident chaperone BiP/HSPA5 to regulate the PERK (EIF2AK3) and IRE1 branches of the UPR. This dual role positions DNAJC3 as a key determinant of cell fate, balancing adaptive translational attenuation via PERK-eIF2??-CHOP signaling against pro-apoptotic outputs. Additional interacting factors include J-proteins and other co-chaperones that fine-tune UPR sensor activity.
In the Jurkat T-cell background, DNAJC3 knockout is anticipated to sensitize cells to ER stress-induced apoptosis by removing the brake on PKR activation and enhancing eIF2?? phosphorylation, leading to sustained translational repression and induction of the pro-apoptotic transcription factor CHOP. This model system also enables the study of how P58IPK loss impacts T-cell receptor-proximal signals, given the known crosstalk between ER stress and immune signaling pathways. The polyclonal nature of the knockout population captures the heterogeneity of gene disruption, providing a more representative picture of the functional consequences compared to monoclonal lines, particularly in pathways with variable expression and compensatory mechanisms.
This DNAJC3 knockout product is suited for a range of biomedical research applications, including detailed UPR pathway analysis, PKR signaling studies, and investigation of translational control mechanisms. Representative assays include Western blotting for phospho-eIF2??, PKR, and DNAJC3, RT-qPCR for CHOP and BiP expression, XBP1 splicing assays, and co-immunoprecipitation of P58IPK with PKR or BiP. Additionally, the cells are compatible with apoptosis assays (Annexin V, caspase-3 activation) under ER stress conditions, phospho-signaling analysis, and flow cytometry-based viability assessments. Applications extend to diabetes research, given DNAJC3’s role in insulin processing, viral host interaction studies, and neurodegenerative disorder modeling where ER stress is a common pathogenic feature. For further information, please contact Ascent Research.
