The EDEM3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This product provides a loss-of-function model for studying the role of EDEM3 in ER-associated degradation (ERAD) and the unfolded protein response (UPR). The polyclonal nature ensures representation of diverse editing events, offering a robust system for investigating EDEM3-dependent processes in a cancer cell context.
The HT29 cell line is a widely used epithelial colorectal adenocarcinoma model originally isolated from a primary tumor of a 44-year-old Caucasian female. HT29 cells exhibit characteristics of intestinal epithelial differentiation and have been extensively employed in cancer biology, signal transduction, and drug discovery research. Their relevance to colorectal cancer makes them a valuable platform for exploring the intersection of ER protein quality control and oncogenesis.
EDEM3 encodes an endoplasmic reticulum (ER) mannosidase that specifically trims ??1,2-mannose residues from misfolded glycoproteins, generating a degradation signal. This trimming facilitates glycoprotein recognition by the ERAD machinery, including the SEL1L-HRD1 complex, derlin, and the AAA-ATPase VCP/p97, leading to retrotranslocation to the cytosol. Downstream, substrates undergo ubiquitination and proteasomal degradation. EDEM3 expression is transcriptionally regulated by ER stress sensors (IRE1, PERK, ATF6) and downstream effectors such as XBP1s and ATF4. Thus, EDEM3 operates at the core of the UPR, linking ER stress detection to the clearance of misfolded proteins.
In HT29 colorectal adenocarcinoma cells, which exhibit altered proteostasis due to oncogenic mutations and rapid proliferation, EDEM3 knockout disrupts the ERAD pathway, potentially causing accumulation of misfolded glycoproteins and chronic ER stress. This may constitutively activate UPR branches such as the IRE1-XBP1 and PERK-ATF4 pathways, influencing cell survival, apoptosis, and sensitivity to chemotherapeutic agents. The HT29 EDEM3 knockout model thus provides a physiologically relevant platform to study how ER protein quality control impacts cancer cell physiology and therapeutic responses.
Researchers can employ these polyclonal knockout cells in a wide range of assays to investigate ER stress and UPR signaling, protein quality control, and cancer biology. For example, western blotting and RT-qPCR can be used to monitor UPR markers (e.g., GRP78/BiP, CHOP) and XBP1 mRNA splicing. Flow cytometry and cell viability assays enable assessment of apoptosis and drug sensitivity. Proteasome activity assays and co-immunoprecipitation allow functional analysis of the ERAD machinery and EDEM3-interacting complexes, while RNA-seq provides a global view of transcriptomic changes. For further inquiries or to request a quote, please contact Ascent Research.
