The IDH2 Knockout CAL-27 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the CAL-27 human tongue squamous cell carcinoma line, engineered for targeted disruption of the IDH2 gene. This polyclonal pool offers a robust loss-of-function model without clonal selection, enabling functional interrogation of IDH2-dependent pathways in a heterogeneous cancer cell context.
The host CAL-27 cell line is an adherent epithelial line established from a human tongue squamous cell carcinoma, retaining wild-type p53 and widely employed as a preclinical model for oral cancer research. Its well-characterized growth properties and genetic background make it a reliable substrate for studying the molecular mechanisms of squamous cell carcinoma pathogenesis, drug responses, and metabolic reprogramming.
IDH2 encodes a mitochondrial NADP+-dependent isocitrate dehydrogenase that catalyzes the oxidative decarboxylation of isocitrate to ??-ketoglutarate (??-KG) and produces NADPH. It functions as a homodimer requiring Mg2? or Mn2? and NADP?, and is regulated by SIRT3-mediated deacetylation and nutrient-sensing networks involving AMPK, mTOR, HIF-1??, and MYC. As a citrate cycle enzyme, IDH2 interfaces with glutamine metabolism via reductive carboxylation, contributing to redox homeostasis, lipid biosynthesis, and anaplerosis. Downstream, ??-KG and NADPH influence ??-KG-dependent dioxygenases such as histone and DNA demethylases, thereby coupling metabolic flux to epigenetic regulation. Representative pathway components include IDH1, ACO2, OGDH, FH, MDH2, GLS, and ACLY.
In the CAL-27 oral squamous cell carcinoma background, IDH2 knockout disrupts mitochondrial citrate cycle flux and diminishes NADPH pools, creating a valuable model to probe metabolic vulnerabilities in head and neck cancers. Given the reliance of many squamous cell carcinomas on glutamine-driven reductive carboxylation and redox buffering, these polyclonal knockout cells enable dissection of how IDH2 loss impacts ??-KG-dependent chromatin modifications, cellular redox status, and sensitivity to metabolic inhibitors or chemotherapeutics. This system is especially suited for exploring TCA cycle rewiring and epigenetic crosstalk in oral tumors, and may support validation of IDH2 as a therapeutic target beyond its established roles in glioma and acute myeloid leukemia.
Typical applications include metabolic flux analyses with Seahorse or isotope tracers to assess TCA cycle rewiring; LC-MS metabolite profiling to measure ??-KG, 2-hydroxyglutarate, and NADP+/NADPH ratios; and epigenomic or transcriptomic profiling via ChIP-seq and RNA-seq. The polyclonal knockout format supports drug target validation, metabolic inhibitor screens, and proliferation or apoptosis studies. These cells provide a consistent IDH2 loss-of-function platform for mechanistic and translational research. For further details, please contact Ascent Research.
