The HEBP1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma cell line HT29, with targeted disruption of the HEBP1 gene. HEBP1 encodes heme-binding protein 1, a critical regulator of intracellular heme availability. The polyclonal format provides a heterogeneous pool of knockout cells, suitable for population-level functional studies without clonal selection biases.
The parental HT29 cell line, isolated from a primary colorectal adenocarcinoma of a 44-year-old female, displays epithelial morphology and enterocyte-like differentiation capacity. Widely used as a model for colorectal adenocarcinoma and intestinal epithelial biology, HT29 cells offer a physiologically relevant system for studying tumorigenesis, metabolic adaptation, and barrier function. Their robust growth and well-characterized genetics facilitate genome editing and downstream functional analyses.
HEBP1 acts as an intracellular heme chaperone, regulating free heme levels for hemoprotein assembly and signaling. It directly binds heme and porphyrins, modulating transcription factors BACH1 and REV-ERB. HEBP1 expression is induced by heme, oxidative stress, and NRF2, forming a feedback loop with iron homeostasis. Downstream, HEBP1 influences heme-dependent enzymes, CLOCK and BMAL1 circadian regulators, and BACH1. This integrates heme metabolism with circadian rhythms, linking to ALAS1, FECH, and HO-1 in the heme pathway.
In HT29 colorectal cancer cells, HEBP1 knockout disrupts heme homeostasis, impairing mitochondrial respiration, redox control, and proliferation. This model enables dissection of heme’s role in tumor cell survival under iron and oxidative stress. Given heme’s link to ferroptosis, these cells are valuable for investigating ferroptotic susceptibility and resistance. Additionally, perturbation of the CLOCK/BMAL1 network by HEBP1 loss provides a platform to study circadian disruption in cancer metabolism.
Applications include western blotting and RT-qPCR for knockout validation and target expression analysis, heme quantification, Seahorse metabolic profiling, and cell proliferation assays. Ferroptosis sensitivity is assessed by lipid peroxidation detection, circadian rhythms by reporter assays, and drug sensitivity testing identifies chemotherapeutic vulnerabilities linked to heme dysregulation. For additional technical information or to discuss custom applications, please contact Ascent Research.
