The IDH2 Knockout LoVo Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population generated from the LoVo human colorectal adenocarcinoma line, designed for loss-of-function studies of the IDH2 gene. This heterogeneous knockout pool results from targeted gene disruption, yielding cells that lack functional IDH2 protein while maintaining the genetic diversity inherent to a polyclonal population, thereby avoiding clonal selection artifacts.
The parental LoVo cell line is an epithelial adenocarcinoma model derived from a metastatic colon tumor, extensively used to investigate colorectal cancer biology, including tumor metabolism, signaling, and drug responses. As adherent cells, LoVo is compatible with high-content imaging, metabolic flux analysis, and standard molecular biology techniques.
IDH2 encodes the mitochondrial NADP+-dependent isocitrate dehydrogenase that converts isocitrate to ??-ketoglutarate (??-KG) and generates NADPH. Its activity is regulated by AMPK, SIRT3, and PPARGC1A, and transcriptionally induced by HIF-1??. IDH2 interacts with citrate synthase, aconitase, and GOT2, using Mg2+ and NADP+. ??-KG is essential for dioxygenases, including TET2 and JmjC histone demethylases, while NADPH maintains redox balance via glutathione reductase. IDH2 loss reduces ??-KG and NADPH, impairing TCA flux, dioxygenase function, and redox homeostasis.
In colorectal adenocarcinoma, IDH2 knockout disrupts the metabolic equilibrium of LoVo cells, which rely on glutaminolysis and oxidative phosphorylation. The decline in NADPH production compromises antioxidant defenses, potentially activating redox-sensitive signaling pathways and creating therapeutic vulnerabilities. Concurrently, lowered ??-KG levels may diminish TET2-mediated DNA demethylation and JmjC-catalyzed histone demethylation, promoting epigenetic alterations that affect proliferation, differentiation, and drug resistance. This polyclonal model therefore offers a valuable system for studying the metabolic-epigenetic interface in colorectal cancer.
These IDH2 knockout cells are suited for metabolic profiling via Seahorse analysis, quantification of NADP+/NADPH ratios and ??-KG levels, and expression studies of TET2, JmjC demethylases, and glutathione reductase by RT?qPCR or Western blotting. Functional assays including cell viability, colony formation, apoptosis, migration, and invasion can assess the impact on tumor aggressiveness and drug sensitivity. The model is also useful for biomarker discovery and exploring resistance mechanisms to metabolism- or epigenetics-targeted therapies. For further details, please contact Ascent Research.
