The BDH2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population designed to disrupt the human BDH2 gene in the HT29 colorectal adenocarcinoma cell line. This loss-of-function model facilitates studies of BDH2’s role in iron homeostasis and ferroptosis regulation without single-cell cloning, preserving the genetic diversity of the parental HT29 cells.
HT29 is a widely used human colorectal adenocarcinoma cell line derived from a primary tumor, serving as an established model for intestinal epithelial biology and colorectal cancer research. These cells exhibit characteristics of differentiated intestinal epithelium and are amenable to studies of colonic tumorigenesis, iron metabolism, and oxidative stress responses.
BDH2 catalyzes the production of the iron-chelating siderophore 2,5-dihydroxybenzoic acid (2,5-DHBA), which modulates the labile iron pool and protects against ferroptosis. The enzyme is regulated by NFE2L2 (NRF2), PPARGC1A (PGC-1??), and iron regulatory proteins IRP1 and IRP2. Downstream, it influences GPX4, ferritin, and TFRC expression, while interacting with mitochondrial proteins SDHA and HSPA9 and iron-sulfur cluster assembly factors ISCU and NFS1. BDH2 disruption abrogates 2,5-DHBA synthesis, enlarging the labile iron pool, elevating lipid peroxidation, and sensitizing cells to ferroptotic death, potentially impairing mitochondrial iron metabolism.
In colorectal cancer, where iron dysregulation and ferroptosis resistance are prevalent, this knockout model enables dissection of iron-dependent cell death pathways. BDH2 ablation in HT29 cells disrupts endogenous iron chelation, revealing vulnerabilities linked to labile iron accumulation and lipid peroxide generation. The model also allows investigation of mitochondrial dysfunction and iron-sulfur cluster biogenesis in a colorectal adenocarcinoma context, informing therapeutic strategies that target ferroptosis or iron metabolism.
This polyclonal knockout cell product is suitable for ferroptosis sensitization assays using erastin or RSL3, lipid peroxidation measurement with C11-BODIPY, and labile iron pool detection via calcein-AM. Additional applications include western blotting for BDH2, GPX4, and ferritin, mitochondrial stress tests with Seahorse analyzers, and co-immunoprecipitation of BDH2-interacting proteins such as SDHA and HSPA9. The model supports RNA-seq transcriptomics and flow cytometric analysis of iron content and ROS. For further details or customized solutions, please contact Ascent Research.
