The DNAJC10 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the DNAJC10 gene in the human HT29 cell line. This loss-of-function model employs CRISPR/Cas9-mediated gene disruption to eliminate functional DNAJC10 protein expression, providing a powerful tool for investigating endoplasmic reticulum (ER) disulfide reductase activity and ER-associated degradation (ERAD). As a polyclonal population, it includes a heterogeneous mix of edited alleles, closely reflecting the genetic diversity encountered in pooled knockout screens and bulk functional assays.
The HT29 host cell line is a well-characterized human colorectal adenocarcinoma epithelial line originally derived from a primary tumor of a female patient. HT29 cells are widely employed as a model for intestinal epithelial differentiation and colorectal cancer, displaying typical epithelial morphology and the capacity to differentiate under specific conditions. They are instrumental in studying cancer biology, ER stress responses, and the interplay between oncogenic signaling and cellular proteostasis mechanisms, making them an appropriate background for examining DNAJC10 function in a tumor-relevant context.
DNAJC10, also known as ERdj5, encodes an ER-resident disulfide reductase that cleaves disulfide bonds of misfolded glycoproteins, priming them for retrotranslocation and proteasomal degradation via the ERAD pathway. It operates downstream of ER stress sensors IRE1??, PERK, and ATF6, and is transcriptionally regulated by XBP1s and ATF4 upon UPR activation. DNAJC10 interacts with key ERAD components including EDEM1, SEL1L, the HRD1 E3 ligase complex, and the chaperone BiP (HSPA5), collectively mediating the recognition and dislocation of terminally misfolded substrates from the ER lumen. Disruption of DNAJC10 impairs this clearance mechanism, resulting in accumulation of misfolded proteins and sustained ER stress signaling.
In the HT29 colorectal cancer model, DNAJC10 knockout holds particular significance for dissecting ERAD-dependent survival mechanisms. Colorectal cancer cells frequently encounter elevated ER stress due to high metabolic demands, secretory activity, and microenvironmental stressors. Loss of DNAJC10 perturbs the balance between ERAD capacity and UPR output, potentially affecting proliferation, apoptosis, and tumorigenicity. This model enables detailed investigation of how ERAD deficiency reshapes stress signaling dynamics in a colorectal adenocarcinoma background, contributing to the understanding of cancer cell adaptation and the identification of synthetic lethal interactions.
This polyclonal knockout cell product is suited for a wide range of research applications, including the study of UPR signaling, ERAD mechanism dissection, and drug screening for ER stress modulators. Representative assays such as Western blotting for UPR markers (BiP, CHOP), RT-qPCR for XBP1 splicing analysis, immunofluorescence for ER stress markers, and co-immunoprecipitation of ERAD complexes (e.g., EDEM1 or SEL1L interactions) can be readily performed. Additionally, viability assays under ER stress induction (e.g., with tunicamycin or thapsigargin) and transcriptomic profiling via RNA-seq provide comprehensive functional readouts. For further technical information, please contact Ascent Research.
