BPGM Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T cells, designed to disrupt the BPGM gene. BPGM (bisphosphoglycerate mutase) encodes the enzyme that catalyzes the isomerization of 1,3-bisphosphoglycerate to 2,3-bisphosphoglycerate, a key step in the Rapoport-Luebering glycolytic shunt. This polyclonal knockout product provides a heterogeneous loss-of-function model for investigating BPGM-dependent metabolic processes and signaling pathways.
HEK293T is a human embryonic kidney cell line stably expressing SV40 large T antigen, known for high transfection efficiency and robust protein expression. It is widely used in biomedical research for viral production, protein overexpression, and gene editing experiments. This epithelial cell line provides a versatile platform for studying gene function in a non-erythroid context.
BPGM is a critical enzyme in 2,3-BPG metabolism, converting 1,3-bisphosphoglycerate to 2,3-BPG, which is an allosteric regulator of hemoglobin oxygen affinity. In addition to its role in erythrocytes, BPGM participates in the glycolytic bypass that modulates levels of glycolytic intermediates. Upstream, BPGM expression is regulated by hypoxia-inducible factor (HIF) and erythropoietin (EPO) signaling, as well as the GATA1 transcription factor. Downstream, BPGM activity influences 2,3-BPG production, hemoglobin oxygen affinity, and glycolytic flux. BPGM interacts with phosphoglycerate kinase (PGK) and bisphosphoglycerate phosphatase, and its activity can indirectly affect hemoglobin function via 2,3-BPG levels.
In HEK293T cells, which lack hemoglobin, the BPGM knockout model is particularly valuable for dissecting the Rapoport-Luebering shunt and its impact on glycolytic metabolism independent of erythroid-specific functions. Loss of BPGM disrupts the conversion of 1,3-BPG to 2,3-BPG, leading to alterations in glycolytic intermediate levels and energy metabolism. This model enables the study of metabolic reprogramming, oxygen sensing, and the HIF pathway without confounding erythroid differentiation. It also provides a system to evaluate how BPGM deletion affects 2,3-BPG synthesis and downstream metabolic pathways.
This polyclonal knockout cell population is suitable for a variety of assays, including Western blotting for BPGM, 2,3-BPG enzymatic assays, LC-MS-based metabolomics, glycolytic flux analysis, oxygen consumption rate (OCR) measurements, and hypoxia response gene expression analyses. Applications encompass studies of 2,3-BPG metabolism, cancer metabolic reprogramming, high-altitude adaptation research, and BPGM inhibitor screening. For further information, please contact Ascent Research.
