The BPGM Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the BPGM gene in the HAP1 cell line. This heterogeneous pool of cells contains targeted disruptions within the BPGM locus, enabling loss-of-function studies without clonal isolation. The polyclonal nature captures a range of editing events, suitable for population-based analyses. This model is a valuable tool for investigating BPGM??s role in metabolism and oxygen sensing.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) line. Its haploid genome simplifies genetic manipulation, often leading to complete loss-of-function phenotypes from single-allele disruptions. HAP1 retains hematopoietic progenitor features and is used for leukemia, signaling, and metabolic studies. Its robust culture and CRISPR compatibility make it ideal for hematological and metabolic gene knockouts.
BPGM encodes bisphosphoglycerate mutase, which converts 1,3-bisphosphoglycerate to 2,3-bisphosphoglycerate (2,3-BPG) in the Rapoport-Luebering shunt of glycolysis. 2,3-BPG is an allosteric effector that binds deoxyhemoglobin, stabilizing its low-oxygen-affinity T-state and promoting oxygen release. BPGM transcription is regulated by erythroid transcription factors GATA1 and KLF1, and is induced by hypoxia-inducible factors. The resultant 2,3-BPG interacts with hemoglobin, modulating oxygen delivery. This pathway integrates BPGM, 1,3-bisphosphoglycerate, 2,3-BPG, and hemoglobin to control oxygen transport.
In the HAP1 hematopoietic background, BPGM knockout allows dissection of the Rapoport-Luebering shunt and glycolytic flux. Although HAP1 cells are leukemic, they provide a genetically clean progenitor-like platform to study BPGM function. Loss of BPGM is predicted to decrease 2,3-BPG, recapitulating aspects of BPGM deficiency linked to hemolytic anemia and erythrocytosis. This model offers insights into red blood cell disorders and hypoxia adaptation.
This polyclonal knockout supports diverse assays: western blot and RT-qPCR for expression analysis, enzyme activity assays for bisphosphoglycerate mutase function, 2,3-BPG quantification, and hemoglobin oxygen dissociation measurements to assess functional impact. Flow cytometry for hematopoietic markers further enables lineage-specific studies. Applications span oxygen homeostasis, high-altitude physiology, and anemia research. For inquiries, contact Ascent Research.
