The EDEM3 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 786-O renal cell carcinoma line. This product provides a heterogeneous pool of cells harboring targeted disruption of the EDEM3 gene, enabling loss-of-function studies without the need for single-cell cloning. The polyclonal format preserves population-level responses and minimizes clonal artifacts, making it suitable for screening applications and pathway interrogation in a cancer cell context.
The parental 786-O cell line is a widely used model of clear cell renal cell carcinoma (ccRCC), characterized by deficiency of the von Hippel-Lindau (VHL) tumor suppressor. VHL loss leads to constitutive activation of hypoxia-inducible factors (HIFs), driving tumorigenesis and metabolic reprogramming. This genetic background renders 786-O cells particularly sensitive to endoplasmic reticulum (ER) stress and proteotoxic insults, as VHL deficiency impinges on adaptive ER quality control mechanisms. Thus, 786-O cells provide a relevant platform to investigate how ER-associated degradation (ERAD) pathways intersect with oncogenic signaling.
EDEM3 encodes an ??-mannosidase that trims Man8GlcNAc2 glycans to Man5-7GlcNAc2 on misfolded glycoproteins in the ER lumen. This step is recognized by the lectins OS9 and XTP3-B, which deliver substrates to the SEL1L-HRD1 dislocation complex for retrotranslocation and proteasomal degradation. EDEM3 acts downstream of UPR sensors IRE1, PERK, and ATF6, and is upregulated by XBP1s under ER stress. It coordinates with ERAD components HERP, Derlin, and VIMP to clear misfolded secretory cargo, maintaining glycoprotein homeostasis.
In the context of VHL-deficient 786-O cells, EDEM3 may play a critical role in managing elevated basal ER stress caused by metabolic dysregulation and secretory demand. Loss of EDEM3 in this background is expected to impair ERAD capacity, leading to accumulation of misfolded glycoproteins and heightened UPR signaling. This knockout model enables dissection of how cancer cells cope with proteotoxic stress and whether EDEM3 dependency represents a vulnerability that can be therapeutically exploited. Moreover, comparing polyclonal EDEM3 knockout pools to parental cells can reveal adaptive responses and off-target compensatory mechanisms that may not emerge in monoclonal lines.
Applications include western blotting for EDEM3 and ERAD markers, RT-qPCR for UPR genes, flow cytometry for ER stress reporters, co-immunoprecipitation with ERAD machinery, and glycoproteomics for substrate identification. Proteasomal degradation assays and cell viability under ER stress (e.g., tunicamycin) assess functional outcomes. The polyclonal format suits phenotypic screens and dose-response studies. For further information or matched parental controls, contact Ascent Research.
