The G6PC3 Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population of human B lymphocytes, engineered to disrupt the G6PC3 gene. This polyclonal format offers a heterogeneous loss-of-function model that captures diverse editing events across the cell pool, enabling robust functional studies without clonal selection bias. The product is designed for researchers investigating glucose metabolism, endoplasmic reticulum (ER) stress, and neutrophil-related disorders in a lymphoblastoid background. It is supplied as a ready-to-use suspension culture, suitable for downstream applications in molecular biology, biochemistry, and drug discovery.
Derived from an EBV-positive Burkitt lymphoma patient, the Raji cell line serves as a well-established, suspension-adapted model for B-cell lymphoma and immune cell biology. These cells retain key characteristics of transformed B lymphocytes, including active immunoglobulin expression and sensitivity to apoptosis-inducing stimuli, making them a versatile platform for studying signaling pathways and metabolic regulation in a cancerous lymphoid context. The parental Raji line is widely used in hematological research, and the introduction of a G6PC3 knockout provides a unique tool to dissect the interplay between glucose homeostasis and lymphocyte survival.
G6PC3 encodes glucose-6-phosphatase catalytic subunit 3, an ER-resident enzyme that hydrolyzes glucose-6-phosphate into glucose and inorganic phosphate, acting in concert with the glucose-6-phosphate transporter SLC37A4 (G6PT). This reaction is critical for maintaining glucose flux across the ER membrane and for mitigating ER stress. The enzyme is transcriptionally regulated by factors such as HIF1A, PPARGC1A, and FoxO1, and is modulated by glucocorticoid receptor signaling. At the molecular level, G6PC3 interacts with ER chaperones including HSPA5 (BiP) and calnexin, and its activity influences the unfolded protein response (UPR) by controlling the level of ER glucose, which impacts protein folding and quality control. Disruption of G6PC3 impairs glucose-6-phosphate hydrolysis, leading to accumulation of phosphorylated glucose, reduced free glucose availability in the ER, and activation of the UPR transducer CHOP, ultimately sensitizing cells to apoptosis through enhanced ER stress signaling.
In the Raji host background, G6PC3 knockout is expected to dysregulate glucose metabolism and amplify ER stress, providing a relevant model for studying neutrophil pathologies such as severe congenital neutropenia type 4 (SCN4), glycogen storage disease type Ib, and Dursun syndrome. Although these diseases primarily affect myeloid lineages, the lymphoblastoid context allows investigation of fundamental mechanisms underlying ER homeostasis and metabolic control that are conserved across cell types. The polyclonal knockout population is particularly suited for experiments that require averaging over multiple genotypes, such as drug screening for compounds that modulate the UPR or restore glucose homeostasis, and for assessing population-level responses to metabolic perturbations.
This knockout product supports a broad array of research applications, including functional studies of glucose-6-phosphatase activity using enzymatic assays, Western blot analysis of G6PC3, BiP, and CHOP expression, and RT-qPCR quantification of UPR target genes. Apoptosis can be measured by flow cytometry with Annexin V staining, while glucose uptake and cell viability assays enable metabolic phenotyping. Luciferase-based UPR reporters further facilitate real-time monitoring of ER stress signaling. The combination of a defined gene disruption in a classic lymphoma line makes this tool valuable for dissecting the role of G6PC3 in lymphocyte biology and for testing therapeutic hypotheses in metabolic and neutropenia-related disorders. For detailed protocols and additional support, please contact Ascent Research.
