The DNAJB12 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa cell line, providing a loss-of-function model for the DNAJB12 gene in a human cervical adenocarcinoma background. This product comprises a heterogeneous pool of cells with targeted disruption of DNAJB12, enabling robust and reproducible studies of ER proteostasis and stress signaling without clonal selection artifacts. CRISPR/Cas9-mediated gene disruption eliminates DNAJB12 co-chaperone activity, making this model suitable for investigating endoplasmic reticulum-associated degradation (ERAD) and the unfolded protein response (UPR) in a well-characterized epithelial cancer cell context.
The host HeLa cell line is an immortalized human cervical adenocarcinoma line with epithelial morphology, widely used as a workhorse in cancer biology, signal transduction, and drug discovery research. Originally isolated from a cervical tumor, HeLa cells exhibit rapid proliferation, strong adhesion, and extensive molecular tool compatibility, including high transfection efficiency and lentiviral transduction. Their well-documented genomic and transcriptomic landscapes facilitate integration with omics approaches, while their transformed phenotype serves as a relevant platform for studying oncogenic signaling and therapeutic vulnerabilities.
DNAJB12 is an ER-resident co-chaperone that functions in recognizing misfolded proteins and targeting them for degradation via the ERAD pathway. It interacts with the major ER chaperone HSPA5/BiP and the AAA-ATPase VCP/p97, forming complexes with ERAD components such as DERL1, SEL1L, and SYVN1 to facilitate retrotranslocation and proteasomal clearance of aberrant polypeptides. These activities are transcriptionally controlled by UPR sensors IRE1, PERK, and ATF6, as well as heat shock factor 1 (HSF1), which collectively orchestrate adaptation to ER stress. Representative pathway members include HSP90B1 and multiple ERAD constituents that cooperate with DNAJB12 to maintain ER homeostasis.
Knockout of DNAJB12 in HeLa cells disrupts ER proteostasis by impairing the recognition and disposal of misfolded ER proteins, leading to their accumulation and chronic activation of the UPR. This perturbation sensitizes cells to proteotoxic insults and may alter the threshold for apoptosis, providing a powerful system to dissect how ER stress interfaces with cancer cell survival, migration, and drug response. The HeLa background offers a well-established model for cervical adenocarcinoma, enabling exploration of DNAJB12??s role in tumor cell adaptation to intrinsic and extrinsic proteotoxic stress.
This knockout cell population is suitable for a broad range of experimental applications, including Western blot analysis of UPR markers such as BiP and CHOP, RT-qPCR profiling of ER stress-responsive genes, immunofluorescence microscopy to assess ER morphology, and flow cytometry-based viability assays under stress conditions. Additionally, the model supports migration and invasion studies, as well as drug sensitivity screens to identify agents that exploit ERAD deficiency. Together, these cells enable detailed mechanistic studies of ER quality control in cancer biology. For further technical information, please contact Ascent Research.
