The EDEM3 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human SK-HEP-1 hepatic adenocarcinoma cell line. This polyclonal product features disruption of EDEM3, encoding an ER degradation-enhancing alpha-mannosidase-like protein critical for endoplasmic reticulum-associated degradation (ERAD). The heterogeneous pool of knockout alleles enables robust loss-of-function studies without clonal selection biases, making it ideal for investigating glycoprotein quality control, ER stress, and liver cancer pathogenesis. CRISPR/Cas9 technology ensures efficient and specific targeting, generating a versatile tool for functional genomics and drug discovery.
SK-HEP-1 is a human hepatic adenocarcinoma cell line isolated from a male patient and widely used as a model for hepatocellular carcinoma (HCC). These cells exhibit epithelial morphology and retain relevant oncogenic pathways, providing a suitable hepatic context for studying ER-associated processes due to the liver’s high secretory activity. Introducing EDEM3 knockout in SK-HEP-1 allows examination of ERAD function in a disease-relevant genetic and phenotypic background.
EDEM3 is an ER-localized alpha-mannosidase-like enzyme that trims mannose residues from misfolded glycoproteins, generating a glycan signal recognized by OS9 and SEL1L. This targets substrates to the HRD1 ubiquitin ligase complex, which includes Derlin-1, for ubiquitination, retrotranslocation by p97/VCP, and 26S proteasomal degradation. EDEM3 expression is regulated by ER stress and the unfolded protein response (UPR), with transcription factors XBP1s, ATF6, and ATF4 controlling its upregulation. Knockout of EDEM3 disrupts this cascade, causing accumulation of misfolded glycoproteins such as mutant alpha-1-antitrypsin Z and eliciting chronic ER stress, thereby positioning EDEM3 as a central proteostasis node.
In SK-HEP-1 liver cancer cells, EDEM3 knockout provides a model to dissect how ERAD deficiency influences HCC biology. Liver cancers often exhibit elevated ER stress due to high metabolic demands; thus, this system enables investigation of adaptive UPR mechanisms and their contribution to tumor survival, proliferation, and drug sensitivity. Additionally, it can model ER stress-related liver pathologies, such as congenital disorders of glycosylation, and facilitate systematic exploration of the interplay between oncogenic signaling and protein quality control.
This polyclonal EDEM3 knockout product supports diverse applications: functional validation of ERAD substrates via western blotting, co-immunoprecipitation, and proteasome activity assays; transcriptomic analyses (RNA-seq, RT-qPCR) to delineate UPR activation; flow cytometry and immunofluorescence for ER stress markers; drug screening for ERAD modulators with MTT/XTT viability and apoptosis assays; and UPR reporter assays. It accelerates research into ERAD mechanisms, liver cancer biology, and proteostasis-targeted therapeutics. For more information, contact Ascent Research.
