DNAJC3 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, engineered to disrupt the DNAJC3 gene. This polyclonal pool provides a heterogeneous loss-of-function model for investigating DNAJC3??s role in endoplasmic reticulum (ER) homeostasis and the unfolded protein response (UPR). The product is suitable for experiments that benefit from a diverse genetic knockout background, enabling robust studies of gene function without clonal isolation.
The parental HeLa cell line originates from a human cervical adenocarcinoma and serves as a widely employed model in cancer biology, cell cycle research, and signal transduction studies. Its epithelial phenotype and ease of genetic manipulation make HeLa cells a tractable system for examining oncogenic pathways and stress responses. The DNAJC3 knockout is introduced into this well-characterized background, preserving the inherent properties of HeLa cells while enabling dissection of ER stress signaling.
DNAJC3 encodes an ER-resident co-chaperone that forms complexes with HSPA5 (BiP) and negatively regulates the ER stress sensor EIF2AK3 (PERK). Under basal conditions, DNAJC3 dampens PERK activity, limiting the phosphorylation of eIF2?? (EIF2S1) and subsequent induction of downstream effectors such as ATF4 and DDIT3 (CHOP). Loss of DNAJC3 disrupts this homeostatic brake, leading to unchecked PERK signaling and heightened sensitivity to ER stress. The protein also interacts with ERN1 (IRE1) and DNAJB11, positioning it as a central modulator of UPR branches. Key pathway components include HSPA5, EIF2AK3, EIF2S1, ATF4, and DDIT3, which collectively govern translation inhibition, protein folding, and apoptosis.
In HeLa cells, which are derived from an adenocarcinoma, the UPR is frequently rewired to support tumor growth and survival. DNAJC3 knockout in this context is expected to sensitize cells to pharmacological ER stress inducers such as thapsigargin and tunicamycin, making the polyclonal population a valuable tool for studying adaptive stress responses relevant to cervical cancer. The heterogeneous knockout background mimics the genetic variability observed in tumor cell populations, providing a more physiologically relevant model compared to clonal lines.
This knockout model supports a range of functional assays, including Western blotting for UPR markers (BiP, p-eIF2??, ATF4, CHOP), RT-qPCR for target gene expression, and cell viability assays under ER stress. It is also amenable to co-immunoprecipitation protocols to probe protein?Cprotein interactions within the chaperone network. Researchers can employ these cells to screen for modulators of the PERK pathway or to investigate the role of co-chaperones in protein folding fidelity. For additional technical details or customized bulk orders, please contact Ascent Research.
