The PHGDH Knockout Raji Polyclonal Cells comprise a population of Raji B lymphoblasts that have undergone CRISPR/Cas9-mediated disruption of the PHGDH gene, generating a heterogeneous pool of loss-of-function alleles. This polyclonal knockout product provides a genetically diverse model system for studying the functional consequences of ablated phosphoglycerate dehydrogenase activity without the clonal selection biases inherent in single-cell-derived lines. The polyclonal format is particularly suited for pooled screening approaches and functional genomics experiments where representation of multiple editing events is advantageous.
The parental Raji cell line is a suspension-adapted human B lymphoblast line established from a Burkitt lymphoma patient. These cells exhibit characteristics of mature B lymphocytes and are widely employed in immunological and cancer research due to their robust growth, well-characterized antigen presentation machinery, and relevance to B-cell malignancies. Raji cells express surface immunoglobulins and MHC molecules, making them a valuable host for investigating the intersection of metabolism and immune function. Their derivation from an aggressive lymphoma also renders them a pertinent model for exploring metabolic dependencies in hematological cancers.
Phosphoglycerate dehydrogenase (PHGDH) catalyzes the first committed step of the serine biosynthesis pathway, converting 3-phosphoglycerate to 3-phosphohydroxypyruvate using NAD+ as a cofactor. This reaction diverts glycolytic intermediates toward the production of serine, which serves as a precursor for glycine, glutathione, nucleotides, and S-adenosylmethionine. PHGDH expression is transcriptionally upregulated by oncogenic MYC, HIF1??, and ATF4, and is further regulated by the PI3K/AKT and NRF2 pathways. It physically interacts with PSAT1, the enzyme catalyzing the subsequent step in serine synthesis, and with the glycolytic enzyme PKM2. The pathway proceeds through PSPH to generate serine, which is then interconverted with glycine by SHMT1 and SHMT2, feeding into one-carbon metabolism via MTHFR and MTR. This network is critical for maintaining redox balance, nucleotide pools, and methylation capacity.
In the context of Raji B lymphoblasts, PHGDH holds particular significance as many lymphomas exhibit a heightened reliance on serine biosynthesis to sustain proliferation and counteract oxidative stress. The knockout of PHGDH in this model allows researchers to dissect the contribution of de novo serine synthesis to lymphoma cell survival, clonal expansion, and metabolic adaptation. Given the role of B lymphocytes in antigen presentation and immune responses, this model also facilitates investigation into how serine pathway activity modulates immune-related functions or sensitivity to therapeutic agents. The polyclonal nature of the knockout population may reveal a spectrum of phenotypic effects reflective of heterogeneous editing outcomes, enriching the analysis of genotype-phenotype relationships.
This polyclonal knockout product is well-suited for a variety of advanced research applications, including metabolic flux analysis using stable isotope-labeled glucose or glutamine tracing to map carbon flow in the absence of PHGDH, LC-MS-based metabolomic profiling of serine, glycine, and downstream metabolites, and cell proliferation and apoptosis assays to assess fitness consequences. It can be employed in synthetic lethality screens with inhibitors of one-carbon metabolism or redox pathways, and in drug sensitivity testing with PHGDH inhibitors such as NCT-503. Additional applications include Western blot verification of pathway disruption, colony formation assays, and Seahorse-based measurements of glycolytic and mitochondrial respiration. For detailed information on lot-specific characteristics and technical support, please contact Ascent Research.
