The EDEM3 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical adenocarcinoma line, engineered to disrupt the EDEM3 gene. This loss-of-function model enables investigation of EDEM3??s role in endoplasmic reticulum-associated degradation (ERAD) and glycoprotein quality control without clonal selection bias. EDEM3 encodes an ??-mannosidase-like protein that trims mannose residues on misfolded glycoproteins to facilitate proteasomal clearance.
HeLa cells are an immortalized cervical cancer line harboring HPV18, widely used for cancer biology and drug discovery. This adherent epithelial line offers robust growth and is well-suited for generating knockout models to study signaling pathways in a malignant context. The cellular background provides relevant oncogenic drivers, enabling detailed investigation of how loss of specific ERAD components affects tumor cell physiology under stress conditions.
EDEM3 acts in the ERAD pathway downstream of UPR sensors IRE1??, PERK, and ATF6. Its expression is induced via XBP1, ATF4, and ATF6 upon ER stress. EDEM3 trims mannose residues from misfolded glycoproteins, generating substrates recognized by OS9 and XTP3-B, which then engage the SEL1L-HRD1 complex for retrotranslocation through Derlin-1. EDEM3 directly interacts with OS9, XTP3-B, SEL1L, HRD1, Derlin-1, and the mannosidase MAN1B1. Knockout impairs ERAD, causing accumulation of misfolded proteins, chronic ER stress, and altered UPR signaling, thus affecting cell survival under proteotoxic conditions.
In HeLa cells, EDEM3 disruption compromises glycoprotein quality control, sensitizing these cancer cells to ER stress-induced apoptosis and death. This creates a model to study how ERAD supports viability and to identify vulnerabilities related to proteasome inhibitor resistance. The HPV18-positive background further permits exploration of viral oncoprotein influence on ER homeostasis, offering insights into cancer-specific stress adaptation mechanisms.
This polyclonal knockout model is applicable for ER stress assays using thapsigargin or tunicamycin, UPR monitoring via qPCR for XBP1 splicing or CHOP, and glycoprotein turnover analysis by cycloheximide chase. Researchers can probe drug resistance by treating with ERAD inhibitors and assess apoptosis by flow cytometry or caspase assays. Additional approaches include GRP78 flow cytometry, ER immunofluorescence, and Western blotting to confirm EDEM3 loss. These cells facilitate detailed dissection of ERAD pathways in cancer and protein misfolding diseases. For inquiries, contact Ascent Research.
