This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1299 human lung carcinoma cell line, in which the EDEM2 gene (ER degradation-enhancing ??-mannosidase-like protein 2) has been disrupted. The polyclonal nature provides a heterogeneous pool of EDEM2-deficient cells, enabling functional studies without clonal selection biases.
The NCI-H1299 host cell line is an immortalized non-small cell lung carcinoma model originating from a lymph node metastasis. It harbors a p53-null background and an activating NRAS Q61K mutation, which drive tumorigenic and metastatic properties. These genetic lesions create a cellular context with high basal proteotoxic stress and dependence on protein quality control mechanisms, making it highly relevant for investigating ER-associated degradation (ERAD) and the unfolded protein response (UPR).
EDEM2 functions as a lectin that recognizes mannose-trimmed N-glycans on misfolded glycoproteins in the ER lumen, acting upstream of the SEL1L-HRD1 E3 ubiquitin ligase complex. It interacts with OS9, XTP3B, and calnexin to direct misfolded substrates toward retrotranslocation and subsequent ubiquitination. The ubiquitinated proteins are extracted by the p97/VCP ATPase and degraded by the proteasome, thereby maintaining ER homeostasis. EDEM2 expression is upregulated by the UPR transcription factors XBP1 and ATF6 under ER stress conditions, linking it directly to adaptive stress signaling.
In the context of NCI-H1299 cells, loss of EDEM2 function is expected to impair clearance of misfolded glycoproteins, leading to accumulation of ERAD substrates, chronic ER stress, and potential activation of apoptotic pathways. Since NSCLC cells often rely on robust ER quality control to survive oncogenic stress, EDEM2 knockout provides a powerful model to explore how disruption of glycoprotein degradation impacts tumor cell fitness, sensitivity to proteasome inhibitors, and the interplay between oncogenic mutations like NRAS Q61K and ER proteostasis.
This polyclonal knockout cell pool is suitable for a variety of applications including analysis of ERAD pathway flux by cycloheximide chase assays, assessment of UPR activation via Western blotting for markers such as BiP and CHOP, and identification of endogenous EDEM2 substrates using proteomics approaches. It can be combined with pharmacological ER stress inducers like tunicamycin or thapsigargin to dissect EDEM2-dependent and independent stress responses. Additionally, cell viability assays and immunofluorescence microscopy can be employed to examine the role of EDEM2 in cancer cell adaptation to proteotoxic insults. For detailed technical specifications or custom inquiries, please contact Ascent Research.
