The BPGM Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-mediated gene-disrupted population derived from the human colorectal adenocarcinoma HT29 cell line. This polyclonal knockout pool carries targeted disruptions in the BPGM gene, leading to loss of bisphosphoglycerate mutase function. The genetically heterogeneous population avoids clonal selection bias, suitable for studying BPGM-dependent processes. The knockout was generated via CRISPR/Cas9 gene editing, resulting in a mixed population of cells harboring diverse genetic alterations at the BPGM locus. This format supports population-level analyses, providing a robust model for investigating BPGM ablation in cancer cells.
The HT29 cell line originates from a primary colorectal adenocarcinoma of a 44-year-old female. These epithelial cells are a well-established colon carcinoma model for studies of tumor progression, differentiation, apoptosis, and drug resistance. HT29 cells retain intestinal epithelial phenotype, making them relevant for colorectal cancer biology. They grow in adherent monolayer and spheroid cultures, supporting various experimental workflows including 3D tumor modeling. Functional glycolysis and hypoxia pathways in HT29 make them ideal for dissecting metabolic adaptation in cancer.
BPGM encodes bisphosphoglycerate mutase, which primarily catalyzes 1,3-BPG to 2,3-BPG conversion. 2,3-BPG allosterically reduces hemoglobin oxygen affinity, promoting oxygen release. In erythroid cells, BPGM is transcriptionally regulated by GATA1 and KLF1. BPGM interacts with glycolytic intermediates and cofactors, linking it to central carbon metabolism. Its product 2,3-BPG influences enzymes such as PGAM1 and LDHA, potentially affecting glycolytic flux. Under hypoxia, HIF1A may modulate BPGM, implicating it in cellular adaptation. Loss of BPGM in HT29 cells disrupts 2,3-BPG synthesis, altering metabolic pathways and hypoxia responses.
Knockout of BPGM in HT29 cells provides a system to study metabolic reprogramming and hypoxic signaling in malignancy. Colorectal tumors show altered glycolysis and oxygen handling; BPGM dysfunction may contribute to metabolic heterogeneity. Eliminating BPGM activity allows investigation of 2,3-BPG depletion effects on glycolytic intermediates, energy metabolism, and cell survival under normoxic and hypoxic conditions. This model is relevant for probing chemoresistance and metabolic symbiosis in the tumor microenvironment. Unlike erythroid models, HT29 cells allow assessment of non-erythroid BPGM functions, clarifying its putative roles in colon carcinogenesis.
This polyclonal knockout cell product is suited for diverse functional studies. Researchers can dissect metabolic roles via Seahorse flux analysis (oxygen consumption, glycolysis) and assess proliferation/apoptosis with MTT and Annexin V assays. Gene expression changes under normoxia/hypoxia are analysable by RT-qPCR or RNA-seq. It serves as a platform for drug target validation and hypoxia-induced gene expression studies. Compatible with 3D spheroid assays, the cells can evaluate differentiation markers or chemotherapeutic responses. Contact Ascent Research for further details.
