IREB2 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population in the HAP1 genetic background, designed for targeted disruption of the IREB2 gene. This knockout model serves as a powerful tool for investigating the cellular roles of iron regulatory protein 2 (IRP2) in post-transcriptional gene regulation and iron homeostasis. The polyclonal nature of this product supplies a heterogeneous pool of edited cells, enabling robust functional studies without clonal selection biases. Researchers can utilize this resource to examine loss-of-function effects on iron-responsive element (IRE)-mediated mRNA regulation in a standardized human cell system.
The HAP1 cell line is a near-haploid human cell line derived from the KBM-7 chronic myelogenous leukemia cell line, exhibiting a predominantly haploid karyotype that simplifies gene-editing approaches. Of male origin, HAP1 cells display an adherent, fibroblast-like morphology and are well-established for genetic knockout studies due to the ease of disrupting single alleles to achieve functional gene inactivation. This background has been extensively employed in functional genomics, allowing efficient generation of knockout models for pathway dissection and high-throughput screening.
IREB2 encodes IRP2, a cytosolic mRNA-binding protein that governs iron homeostasis by binding to IRE motifs in target mRNAs. Under iron deficiency, IRP2 stabilizes TFRC mRNA and represses translation of FTH1, FTL, and SLC40A1, thereby increasing iron uptake and decreasing storage and export. IRP2 is degraded by the E3 ubiquitin ligase FBXL5 in iron-replete conditions, and its stability is also influenced by hypoxia through HIF-1??. Downstream targets include ALAS2 and HIF-2??, linking iron regulation to erythropoiesis and oxygen sensing. IRP2 interacts with the SKP1-CUL1-F-box complex and IRE motifs, and feeds into ferroptosis and HIF-1 signaling pathways.
In the HAP1 model, disruption of IREB2 provides a simplified platform to dissect IRP2-dependent regulatory networks, as the near-haploid genome allows straightforward loss-of-function phenotypes. This polyclonal knockout pool is particularly advantageous for examining heterogeneous responses to iron stress, avoiding artifacts that may arise from clonal expansion. Originating from a chronic myelogenous leukemia background, HAP1 cells offer a relevant context for exploring iron dysregulation in cancer, especially given the emerging role of ferroptosis in tumor biology. The combination of IRP2 loss in a haploid system enables clear readouts in assays monitoring intracellular labile iron pool dynamics, transferrin receptor expression, and ferritin levels, offering insight into both basal iron management and stress-induced adaptations.
Applications span dissecting iron homeostasis, ferroptosis mechanisms, and cancer metabolism. Typical assays include western blotting for IRP2, TFRC, and ferritins; RT-qPCR for IRE-containing transcripts; 55Fe uptake/efflux assays; ferroptosis induction and viability measurements; and co-immunoprecipitation with FBXL5. Immunofluorescence and RNA immunoprecipitation enable further characterization. The model supports drug target validation for neurodegenerative disorders and genetic interaction screens. For further information, please contact Ascent Research.
