The NGLY1 Knockout Raji Polyclonal Cells product comprises a polyclonal population of Raji B lymphoblastoid cells engineered via CRISPR/Cas9-mediated disruption of the human NGLY1 gene. This knockout model provides a heterogeneous pool of cells with loss-of-function mutations across the NGLY1 locus, enabling robust functional studies without clonal bias. The polyclonal format captures the spectrum of editing outcomes, making it suitable for population-level analyses of N-glycanase 1 deficiency and its downstream effects.
The Raji host cell line is a well-characterized B lymphoblastoid model derived from a Burkitt’s lymphoma patient, stably carrying Epstein-Barr virus (EBV) and expressing B cell lineage markers including CD19, CD20, and surface IgM. This cell line is extensively used for investigating B cell biology, lymphomagenesis, and EBV-driven oncogenic mechanisms. Its transformed phenotype and active protein synthesis machinery render it particularly relevant for examining protein quality control and proteotoxic stress responses in a cancer context.
NGLY1 encodes peptide:N-glycanase, a cytosolic enzyme that deglycosylates misfolded N-linked glycoproteins retrotranslocated from the ER during ERAD. It facilitates proteasomal degradation of these substrates and proteolytically activates transcription factor NFE2L1/NRF1, a master regulator of proteasome biogenesis. NGLY1 operates within a network including p97/VCP, HRD1-SEL1L E3 ligase, and RAD23A/B, responding to upstream ER stress sensors (ATF6, IRE1??, PERK) and driving proteasome subunit gene expression (PSMA, PSMB, PSMC). Components like Derlin and BAG6 assist in substrate delivery.
In the Raji Burkitt’s lymphoma background, NGLY1 disruption dissects ERAD and proteostasis in B cell transformation and drug sensitivity. Cancer cells often rely on efficient protein degradation; loss of NGLY1 impairs glycoprotein clearance and blunts NFE2L1/NRF1-mediated proteasome induction, potentially sensitizing cells to proteasome inhibitors and ER stress agents. This model recapitulates NGLY1 deficiency features, including neurodegeneration, liver dysfunction, and intellectual disability, in a proliferating B cell environment for tissue-specific studies. The EBV-positive status adds complexity, as viral pathways may intersect with ERAD, enabling exploration of viral oncoprotein handling.
Applications include Western blot analysis of NGLY1 and ERAD substrates, RT-qPCR of proteasome genes, NRF1 reporter assays, and drug sensitivity testing with proteasome inhibitors like bortezomib. Apoptosis and viability can be assessed by flow cytometry, while immunofluorescence visualizes ER stress markers. These cells support NGLY1 deficiency disease modeling, proteostasis research in lymphomas, and validation of ERAD-ubiquitin-proteasome targets. For product details or technical inquiries, contact Ascent Research.
