The EDEM3 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Jurkat T lymphocytes harboring targeted disruption of the EDEM3 gene, establishing a loss-of-function model to dissect endoplasmic reticulum-associated degradation (ERAD) pathways. This knockout product is supplied as a heterogeneous pool of edited cells, enabling functional studies without clonal selection, and is suited for investigating glycoprotein quality control and ER stress responses in a human T-cell context.
Jurkat cells are an immortalized human T-lymphocyte line originally derived from the peripheral blood of a 14-year-old male with acute lymphoblastic leukemia (ALL). This well-characterized model recapitulates key aspects of T-cell biology and leukemogenesis, including rapid proliferation, expression of T-cell surface markers, and robust signaling through pathways governing survival and apoptosis. Its hematopoietic origin and sensitivity to ER perturbations make it an ideal host for examining how ERAD disruption affects lymphocyte homeostasis and malignancy.
EDEM3 encodes an ER-resident mannosidase that accelerates ERAD by trimming mannose residues from misfolded glycoproteins, thereby committing them to retrotranslocation and proteasomal destruction. Its expression is strongly induced under ER stress downstream of the IRE1??-XBP1s, ATF6, and PERK-eIF2??-ATF4 signaling arms of the unfolded protein response (UPR). EDEM3 interacts with key ERAD machinery components??including SEL1L, HRD1, OS9, XTP3-B, calnexin, and calreticulin??and forms functional complexes with the retrotranslocation channel (Derlin-1, VIMP) and the segregase p97/VCP. Thus, EDEM3 serves as a critical checkpoint that couples glycoprotein misfolding recognition to ubiquitin-proteasome-dependent clearance.
In Jurkat T cells, loss of EDEM3 disrupts ERAD, leading to accumulation of aberrant glycoproteins, chronic ER stress, and potential activation of death signaling pathways. This model enables precise interrogation of how ER homeostasis maintains T-cell viability and how its collapse contributes to leukemic cell sensitivity or resistance to proteotoxic insults. Because EDEM3 is regulated by major UPR branches, its knockout can reveal branch-specific contributions to ER stress outcomes and identify vulnerabilities that may be exploited therapeutically in T-cell malignancies.
Typical experimental applications include profiling UPR target gene expression (e.g., BiP, CHOP) by RT-qPCR or western blotting, measuring apoptosis via Annexin V flow cytometry, assessing proteasomal activity, performing cycloheximide chase assays to monitor glycoprotein turnover, and conducting transcriptome-wide analyses under chemical ER stressors. The model is also valuable for screening small molecules that modulate ERAD or alter leukemic cell fitness. For further information, please contact Ascent Research.
