The IDH1 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the IDH1 gene in the human HT29 colorectal adenocarcinoma cell line. This loss-of-function model enables systematic investigation of IDH1-dependent metabolic and signaling processes without relying on specific editing outcomes at the target locus. The polyclonal format reflects a heterogeneous pool of edited cells, providing a robust system for studying gene function in a population context.
HT29 cells are derived from a human colorectal adenocarcinoma and exhibit an adherent epithelial morphology. They serve as a well-established model for colon epithelial biology and colorectal cancer research. The HT29 line retains multiple cancer-relevant features, including the ability to form colonies and respond to metabolic perturbations, making it a suitable background for assessing oncogenic mechanisms and therapeutic vulnerabilities.
IDH1 encodes cytosolic isocitrate dehydrogenase 1, which catalyzes the oxidative decarboxylation of isocitrate to ??-ketoglutarate (??-KG) with the concomitant reduction of NADP+ to NADPH. This reaction is a key node connecting the citrate cycle, glutamine metabolism, and cellular redox homeostasis. IDH1 activity is regulated by upstream factors including HIF-1??, the PI3K/AKT pathway, MYC, and glucose availability. IDH1 functions as a homodimer and operates in parallel with mitochondrial IDH2. The NADPH produced by IDH1 supports lipid biosynthesis and maintains antioxidant defenses, while ??-KG serves as a co-substrate for dioxygenases such as TET2 and PHD2, which regulate DNA methylation and hypoxia-inducible factor stability, respectively. Disruption of IDH1 therefore diminishes cytosolic NADPH pools, sensitizes cells to oxidative stress, impairs lipid synthesis, and alters the activity of ??-KG-dependent epigenetic modifiers and oxygen sensors.
In the context of HT29 colorectal cancer cells, IDH1 knockout provides a powerful tool to assess the contribution of cytosolic isocitrate metabolism to tumor cell fitness, redox adaptation, and metabolic plasticity. The loss of IDH1 may compromise the cell’s ability to counteract oxidative stress and could disrupt biosynthetic pathways required for rapid proliferation. This model is particularly relevant for exploring synthetic lethal interactions and for testing inhibitors targeting compensatory metabolic routes. Because HT29 cells carry oncogenic mutations, the interplay between IDH1 loss and background genetic alterations can be directly examined.
Researchers can apply the IDH1 Knockout HT29 Polyclonal Cells in diverse experimental workflows, including metabolic flux analysis using Seahorse analyzers or isotopic tracing, quantification of ??-KG and NADPH/NADP+ ratios, and ROS detection assays. The model supports drug sensitivity screening, synthetic lethality studies, and epigenetic profiling via RNA-seq and DNA methylation analysis. Functional assays such as cell proliferation, colony formation, and apoptosis measurements can characterize the phenotypic consequences of IDH1 disruption. For further information, please contact Ascent Research.
