The IDH2 knockout Ca Ski polyclonal cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the mitochondrial isocitrate dehydrogenase 2 (IDH2) gene in the human Ca Ski cervical carcinoma cell line. This product provides a genetically heterogeneous loss-of-function model, enabling robust investigation of IDH2-dependent metabolic and epigenetic functions while avoiding biases introduced by single-cell cloning.
Ca Ski cells are a human female-derived epithelial cell line originating from a cervical squamous cell carcinoma metastasis. These cells stably harbor human papillomavirus type 16 (HPV16) and serve as an established model for studying HPV-driven cervical carcinogenesis. Their transformed phenotype and epithelial origin render them highly relevant for examining the interplay between viral oncogenesis and host cell metabolism.
IDH2 encodes a mitochondrial NADP+-dependent isocitrate dehydrogenase that catalyzes the conversion of isocitrate to ??-ketoglutarate (??-KG) with concomitant NADPH production. IDH2 activity is regulated by substrate availability (isocitrate, NADP+) and SIRT3-mediated deacetylation. The product ??-KG acts as a critical co-substrate for TET DNA demethylases and JmjC-domain histone demethylases, while NADPH is essential for mitochondrial redox balance and anabolic pathways. IDH2 also interacts with mitochondrial complex I and contributes to the TCA cycle. CRISPR/Cas9-mediated disruption of IDH2 reduces ??-KG and NADPH levels, impairing ??-KG-dependent dioxygenase function and perturbing DNA and histone methylation patterns, thereby affecting cellular epigenetics and redox homeostasis.
In the HPV16-positive Ca Ski background, IDH2 knockout serves as a valuable tool for dissecting metabolic dependencies in cervical cancer. HPV oncoproteins E6 and E7 drive proliferation and alter metabolic pathways; loss of IDH2 may uncover specific vulnerabilities in TCA cycle flux and NADPH-dependent antioxidant defenses. Moreover, because HPV-associated cancers exhibit profound epigenetic alterations, this model enables investigation of how IDH2-derived ??-KG modulates the activity of TET and JmjC enzymes, with potential impacts on gene expression programs. The polyclonal nature of the knockout population captures a range of IDH2 disruption efficiencies, more faithfully reflecting the heterogeneity of tumor cell responses.
Researchers can utilize this polyclonal knockout model in a suite of functional assays. IDH2 gene disruption can be validated by western blotting and RT-qPCR. Downstream metabolic consequences are measurable via intracellular ??-KG quantification and NADP+/NADPH ratio assays. Cancer-relevant phenotypes, including proliferation, colony formation, and migration/invasion, can be assessed alongside DNA methylation analyses to probe epigenetic changes. Drug sensitivity studies may reveal altered responses to therapeutics targeting metabolic or epigenetic pathways. This product is well-suited for investigations in cancer metabolism, TCA cycle dysfunction, redox biology, and HPV-related cervical cancer. For inquiries, contact Ascent Research.
