PHGDH Knockout PK-15 Cell Line

The choice depends on whether you are studying PHGDH (phosphoglycerate dehydrogenase)'s role as the rate-limiting enzyme of de novo serine biosynthesis or modeling its applications in cancer metabolism and porcine biology. The Knockout line is the standard tool for asking whether PHGDH is required for these processes — PHGDH catalyzes the first and rate-limiting step of the serine synthesis pathway (SSP), oxidizing 3-phosphoglycerate (from glycolysis) to 3-phosphohydroxypyruvate using NAD⁺ as cofactor; downstream PSAT1 and PSPH complete serine biosynthesis. Serine is a major one-carbon donor (through SHMT1/2-mediated conversion to glycine and methylenetetrahydrofolate), supporting nucleotide synthesis, methylation reactions, redox homeostasis (GSH synthesis), and lipid biosynthesis. Overexpression is useful for studying PHGDH gain-of-function effects. For cancer metabolism research, the EDITGENE PHGDH Knockout in PK-15 is uniquely valuable — PHGDH genomic amplification has been characterized in ⭐⭐ basal-like/triple-negative breast cancer (~6%) and melanoma, where it drives serine auxotrophy escape and is an established cancer metabolism target; PHGDH is a leading 'metabolic oncogene' candidate. Rescue with wild-type or catalytically-dead PHGDH enables structure-function studies. The knockout is a critical specificity tool for ⭐ PHGDH-selective inhibitors (NCT-503, CBR-5884, WQ-2101, and emerging compounds in preclinical/clinical development), serine deprivation cancer therapy research, and emerging PHGDH-related porcine biology (PHGDH is also important for skeletal muscle and embryonic development). PK-15-specific considerations: • PK-15 (CCL-33) is a porcine (Sus scrofa) kidney epithelial cell line widely used in veterinary virology as a primary host for porcine circovirus (PCV2), classical swine fever virus (CSFV), pseudorabies virus (PRV), porcine parvovirus, and other swine pathogens. • Lentiviral transduction is supported with moderate efficiency, but optimization may be required compared to standard human immortalized lines. • PK-15 cells contain endogenous porcine type C retrovirus (PERV) particles and are reported to be persistently positive for PCV2 antigens — these features should be considered when designing viral immunology and xenotransplantation-relevant experiments. • PK-15 cells retain intact type I IFN signaling (unlike Vero cells), making them suitable for innate immunity studies but potentially limiting for some virus isolation workflows. • PK-15 cell clones with differing virus permissivity have been characterized — phenotypic heterogeneity should be assessed before key experiments.

Primary applications: • Serine biosynthesis: 3-phosphoglycerate → serine flux analysis by 13C-glucose tracing in PHGDH-null cells — abolished given PHGDH's rate-limiting role. • Serine auxotrophy: serine-free media growth analysis — PHGDH-null cells should become serine-auxotrophic. • One-carbon metabolism: serine-derived methylation, nucleotide synthesis, and GSH levels analysis. • PHGDH inhibitor specificity: critical genetic control for ⭐ NCT-503, CBR-5884, WQ-2101 in cancer drug development. • Cancer metabolism modeling: in heterologous PHGDH-amplified cancer-relevant contexts, serine synthesis pathway dependence studies. • Porcine biology: in PK-15 porcine kidney context, characterization of PHGDH in swine cell metabolism for veterinary/comparative biology research. EDITGENE recommends this porcine PK-15-based model for researchers investigating serine metabolism, cancer metabolism, and emerging PHGDH-targeted therapeutic development with a unique veterinary/comparative biology dimension.

Yes. PHGDH rescue experiments are well-established for serine biosynthesis research: • Construct design: use a codon-modified PHGDH sequence with a small C-terminal tag (FLAG, HA). PHGDH has the canonical D-2-hydroxyacid dehydrogenase architecture with N-terminal substrate-binding domain, central NAD-binding Rossmann fold, and C-terminal ACT regulatory domain (serine feedback inhibition site) — preserve all elements. • Catalytically-dead rescue: active site mutations (catalytic His or Glu) abolish PHGDH activity. • Feedback-resistant rescue: ACT domain mutations disrupt serine feedback inhibition, generating constitutively active PHGDH. • Species considerations: porcine PHGDH (NCBI ID 100144529) and human PHGDH share substantial homology; either species cDNA can be used for rescue with appropriate species-matching for in vivo veterinary studies. • Functional readout: rescue should restore serine prototrophy and 13C-glucose-traced serine synthesis. PK-15-specific considerations: • PK-15 (CCL-33) is a porcine (Sus scrofa) kidney epithelial cell line widely used in veterinary virology as a primary host for porcine circovirus (PCV2), classical swine fever virus (CSFV), pseudorabies virus (PRV), porcine parvovirus, and other swine pathogens. • Lentiviral transduction is supported with moderate efficiency, but optimization may be required compared to standard human immortalized lines. • PK-15 cells contain endogenous porcine type C retrovirus (PERV) particles and are reported to be persistently positive for PCV2 antigens — these features should be considered when designing viral immunology and xenotransplantation-relevant experiments. • PK-15 cells retain intact type I IFN signaling (unlike Vero cells), making them suitable for innate immunity studies but potentially limiting for some virus isolation workflows. • PK-15 cell clones with differing virus permissivity have been characterized — phenotypic heterogeneity should be assessed before key experiments.