The ATF6B Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HEK293T human embryonic kidney cell line, offering a loss-of-function model for ATF6B, a negative regulator of the unfolded protein response (UPR). This polyclonal pool contains diverse ATF6B-disrupted cells, enabling unbiased functional studies of ER stress signaling without clonal selection artifacts. Designed for applications in ER stress biology, cancer research, and neurodegenerative disease modeling, these cells provide a versatile tool for dissecting ATF6B-mediated signaling and its impact on cellular homeostasis.
The HEK293T host line originated from HEK293 cells and stably expresses SV40 large T antigen, facilitating episomal plasmid replication and high transfection efficiency. As adherent epithelial cells, HEK293T are widely employed for protein expression, viral production, and gene function analyses. Their robust growth and well-characterized ER stress responses make them ideal for UPR research.
ATF6B functions as an ER stress-sensing bZIP transcription factor that represses ATF6A target genes, thereby attenuating the UPR. Under stress, BiP/GRP78 dissociation permits ATF6B trafficking to the Golgi, where cleavage by S1P and S2P proteases releases a nuclear fragment that heterodimerizes with ATF6A or NF-Y to tune expression of XBP1, HSPA5/BiP, DDIT3/CHOP, and ER chaperones. This transcriptional repression is crucial for cellular adaptation to ER stress and for preventing excessive UPR activation. It operates alongside IRE1, PERK, and eIF2?? pathways, integrating signals to balance survival and apoptosis.
In HEK293T cells, ATF6B knockout relieves repression of ATF6A-driven transcription, potentially hyperactivating UPR outputs and altering stress responses. This model enables detailed analysis of how ATF6B-mediated negative feedback controls UPR threshold and duration, with relevance to cancer, neurodegeneration, and metabolic disease. Researchers can evaluate changes in cell viability, apoptosis, and downstream effector expression upon challenge with tunicamycin or thapsigargin.
The ATF6B Knockout HEK293T Polyclonal Cells are applicable to a broad set of experimental workflows, including quantitative RT-qPCR and Western blotting to monitor UPR target gene and protein expression, XBP1 splicing assays to assess IRE1 activity, and immunofluorescence to track ATF6B localization. They also provide a robust cellular substrate for high-throughput screening of ER stress pathway modulators, facilitating drug discovery efforts in oncology, metabolic disorders, and neurodegeneration. For detailed protocols and technical assistance, please contact Ascent Research.
