The DNAJB2 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat T-lymphocyte cell line. This product provides a loss-of-function model for the DNAJB2 gene, which encodes a co-chaperone of the HSP70 family. The polyclonal nature of the knockout ensures a heterogeneous population of edited cells, enabling robust representation of gene disruption effects across the cellular pool. The gene disruption was introduced via CRISPR/Cas9-mediated genome editing, without selection for monoclonal clones, thereby preserving the diversity of knockout alleles. This format is suitable for researchers investigating the functional consequences of DNAJB2 ablation in a human T-cell background.
The host cell line, Jurkat, is an immortalized human T lymphocyte originally isolated from a patient with acute T cell leukemia. This cell line has been extensively utilized in immunology, signal transduction, and cancer biology research. Jurkat cells exhibit rapid proliferation and a relatively stable karyotype, making them a convenient model for gene perturbation studies. They express core components of the cellular stress response machinery, including heat shock proteins and the ubiquitin-proteasome system. Consequently, Jurkat cells provide a physiologically relevant environment for examining the role of DNAJB2 in proteostasis and stress signaling within lymphocytes.
DNAJB2 is a member of the DNAJ/HSP40 family of co-chaperones and functions as an obligate partner of HSP70 chaperone proteins. Mechanistically, DNAJB2 binds to client proteins and delivers them to HSP70 homologs such as HSPA1A and HSPA8, facilitating ATP-dependent folding or, in conjunction with the E3 ubiquitin ligase STUB1/CHIP, targeting misfolded substrates for ubiquitin-dependent proteasomal degradation. The activity of DNAJB2 is regulated upstream by the transcription factor HSF1, which is activated under conditions of heat shock, oxidative stress, or endoplasmic reticulum stress. Loss of DNAJB2 disrupts these interactions, leading to impaired HSP70-mediated substrate processing and reduced proteasomal turnover, ultimately resulting in the accumulation of aggregated proteins and autophagy induction. Key pathway components include HSPA1A, HSPA8, STUB1, and the proteasomal subunit PSMC1.
In the Jurkat T-cell context, DNAJB2 knockout provides a powerful tool for dissecting the cellular response to proteotoxic stress. T lymphocytes are particularly sensitive to disruptions in protein folding quality control due to their high metabolic demands and exposure to oxidative stress during immune activation. By eliminating DNAJB2, this model can recapitulate aspects of Charcot-Marie-Tooth disease and distal hereditary motor neuropathy, disorders linked to dysfunctional protein quality control in neurons and potentially glial-immune interactions. Moreover, it enables the investigation of how impaired chaperone networks affect T-cell viability, cytokine production, and stress-induced apoptosis. This polyclonal knockout population thus serves as a bridge between fundamental cell biology and translational research into neurodegenerative conditions.
Researchers can employ this knockout model in a diverse array of experimental applications. Protein aggregation can be monitored via Western blotting and detergent-insolubility fractionation, while flow cytometry allows assessment of stress-induced apoptosis using markers such as annexin V. RT-qPCR can quantify transcriptional changes in heat shock response genes downstream of HSF1. Co-immunoprecipitation experiments can verify the loss of DNAJB2 interaction with HSPA1A/HSPA8, and ubiquitination assays combined with proteasome activity measurements can delineate shifts in degradation pathways. Additionally, this model is suitable for investigating autophagy cargo targeting and the crosstalk between the ubiquitin-proteasome system and autophagic clearance. For further technical details or custom orders, please contact Ascent Research.
