IDH1 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the IDH1 gene in the human HAP1 cell line. This heterogeneous pool provides a robust loss-of-function model for studying cytosolic isocitrate dehydrogenase 1 biology, ensuring comprehensive gene inactivation across the culture while preserving population-level diversity and eliminating the need for clonal selection.
HAP1 is a near-haploid adherent cell line derived from the male KBM-7 chronic myeloid leukemia isolate, expressing the BCR-ABL1 fusion. Its haploid nature simplifies knockout generation by allowing single-allele targeting, and it retains hematopoietic progenitor features, serving as a versatile platform for functional genomics and cancer metabolism studies without the confounding effects of a full diploid genome.
IDH1 encodes an NADP+-dependent enzyme that homodimerizes and uses Mg2?/Mn2? cofactors to convert isocitrate to ??-ketoglutarate (??-KG) and NADPH. This activity sits at the nexus of the TCA cycle, lipid biosynthesis, and redox homeostasis. IDH1 is transcriptionally regulated by HIF1A and MYC and functionally collaborates with IDH2, GLS1, and GLUD1. The ??-KG product serves as a co-substrate for TET2 DNA demethylase and JmjC histone demethylases (e.g., KDM4A), linking metabolism to epigenetic control, while NADPH drives reductive biosynthesis (e.g., FASN) and antioxidant defense through glutathione reductase (GSR). Thus, IDH1 knockout disrupts both metabolic flux and epigenetic maintenance.
In the BCR-ABL1-driven HAP1 background, IDH1 loss unmasks dependency on glutaminolytic ??-KG production and heightens vulnerability to oxidative stress, reflecting metabolic rewiring observed in IDH1 wild-type leukemias and gliomas. This model is ideal for dissecting how cytosolic NADPH generation buffers redox perturbations and influences chemosensitivity in transformed hematopoietic cells.
Typical applications include metabolic flux analysis, ??-KG/NADPH quantification, lipidomics, ROS assays, and chemosensitivity screening. Western blotting and RT-qPCR confirm knockout, while chromatin profiling and RNA-seq reveal epigenetic and transcriptional consequences. These polyclonal cells support drug target validation and functional genomics investigations. For additional information or technical support, contact Ascent Research.
