The DNAJB14 Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-mediated gene-edited polyclonal cell population derived from the Jurkat T-lymphocyte cell line, designed for the disruption of the DNAJB14 gene. This product provides a heterogeneous pool of cells with targeted loss of DNAJB14 function, enabling robust investigation of its biological roles in a human T-cell background without the need for clonal isolation. Researchers can utilize these cells to study the consequences of DNAJB14 ablation on nonsense-mediated mRNA decay and related processes.
The Jurkat cell line, originally established from the peripheral blood of a 14-year-old male patient with acute T-cell leukemia (T-ALL), is an immortalized T-lymphocyte model extensively employed for studying T-cell receptor (TCR) signaling, apoptosis, and leukemogenesis. These cells exhibit characteristic T-cell markers and retain key signaling pathways, providing a well-defined experimental platform for functional genomics and cancer biology research. Their rapid growth and stable phenotype facilitate reproducible loss-of-function analyses.
DNAJB14 encodes a DnaJ family co-chaperone that recruits heat shock protein HSPA8/HSC70 to the ribosome, where it facilitates nonsense-mediated mRNA decay (NMD) via interaction with the core NMD factor UPF1. This process degrades transcripts with premature termination codons, requiring the coordinated action of SMG6, SMG7, eRF1, and eRF3. Transcription of DNAJB14 is activated by stress-sensitive transcription factors HSF1 and ATF4, linking the heat shock response to RNA quality control. Consequently, DNAJB14 plays a central role in preventing the expression of aberrant proteins and maintaining cellular homeostasis.
In Jurkat T-lymphocytes, knockout of DNAJB14 disrupts NMD, leading to stabilization of mRNAs with premature stop codons and potential accumulation of truncated proteins. This may provoke cellular stress responses and alter T-cell receptor signaling dynamics, as NMD is known to modulate the expression of genes involved in apoptosis and stress adaptation. Given the involvement of NMD in tumor suppression and stress pathways, this model is particularly valuable for dissecting the interplay between RNA quality control and T-cell biology in the context of leukemia and other cancers.
The DNAJB14 Knockout Jurkat Polyclonal Cells are suitable for a wide array of downstream applications. Researchers can perform RNA sequencing to characterize transcriptome-wide shifts in gene expression, RT-qPCR and Western blotting to quantify NMD substrates, and co-immunoprecipitation to assess interactions with UPF1 or ribosomal components. Additionally, flow cytometry-based assessment of T-cell activation markers and apoptosis assays under stress conditions provide functional readouts of NMD impairment. For further product information, please contact Ascent Research.
