The HEBP1 Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the HEBP1 gene. This product provides a heterogeneous mixture of edited HAP1 cells carrying targeted disruptions in the HEBP1 locus, generated via CRISPR/Cas9-mediated gene disruption. Unlike clonal cell lines, the polyclonal format preserves population-level diversity, which is advantageous for studying gene function without clonal selection biases. The knockout model serves as a powerful tool for dissecting the contributions of HEBP1 to intracellular heme dynamics and oxidative stress management.
HAP1 is a near-haploid human cell line derived from the chronic myeloid leukemia (CML) cell line KBM-7. Its haploid karyotype simplifies genetic analysis, as only one allele needs to be disrupted to achieve a functional knockout. HAP1 cells retain leukemic hematopoietic characteristics, making them a relevant model for hematological research. The cell line grows adherently and is amenable to various genetic perturbations, supporting a wide range of functional assays. This background is particularly useful for studying genes involved in processes such as heme metabolism and oxidative stress, where the interplay between heme availability and hematopoietic cell function is critical.
HEBP1 encodes an intracellular heme-binding protein that buffers free heme, regulating its availability for hemoproteins and preventing heme-catalyzed ROS generation. The protein binds heme and porphyrins and forms homodimers. Its expression is induced by heme and controlled by transcription factors NRF2 and GATA1. By controlling labile heme, HEBP1 influences the Bach1/NRF2 oxidative stress response: high heme promotes NRF2 activation and Bach1 degradation, while low heme allows Bach1-mediated repression of antioxidant genes. HEBP1 thus functionally intersects with heme metabolism components HMOX1, FECH, ALAS1, and SLC48A1, integrating heme availability with cellular redox balance.
In the context of HAP1 CML-derived cells, HEBP1 knockout provides a physiologically relevant platform to investigate heme homeostasis in leukemia biology. Leukemic cells often exhibit altered oxidative stress responses and heme metabolism, which can influence proliferation and drug resistance. Disruption of HEBP1 in this background may perturb heme availability, leading to increased oxidative stress and altered signaling through the Bach1/NRF2 pathway. This model enables the study of how heme-dependent pathways affect leukemic cell survival and response to therapies that induce oxidative stress, such as certain chemotherapeutics or heme-targeting agents. The haploid nature of HAP1 cells ensures robust genotype-phenotype correlations, enhancing the utility of the model for mechanistic studies.
These cells are suitable for heme trafficking studies, functional genomics screens, and oxidative stress pathway analysis. Supported assays include Western blotting, RT-qPCR, heme quantification, ROS detection with H2DCFDA, viability under heme stress, iron assays, and Bach1/NRF2 luciferase reporter assays. They also enable drug sensitivity screens for heme-related cancer pathways. For further information, please contact Ascent Research.
