The ADPGK Knockout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population derived from the human colorectal adenocarcinoma HT29 cell line, featuring disruption of the ADPGK gene. This polyclonal format provides a heterogeneous pool of cells carrying diverse ADPGK gene edits, enabling robust loss-of-function analysis without the clonal selection bias associated with single-cell-derived lines. The CRISPR/Cas9-mediated gene disruption serves as a powerful tool for interrogating ADPGK function in colorectal cancer metabolism and hypoxia response pathways.
HT29 cells are an established human colorectal adenocarcinoma line originally isolated from a primary tumor of a 44-year-old Caucasian female. These cells exhibit an epithelial morphology and are extensively utilized as an intestinal epithelial model in cancer research. The HT29 background is particularly relevant for studying colorectal tumor biology, as these cells retain key characteristics of colon adenocarcinoma, including rapid proliferation and the capacity for metabolic adaptation under various microenvironmental conditions.
The ADPGK gene encodes an ADP-dependent glucokinase that catalyzes the phosphorylation of glucose to glucose-6-phosphate using ADP rather than ATP. This enzyme is critical for sustaining glycolytic flux under ATP-limiting conditions, such as hypoxia. ADPGK is transcriptionally regulated by hypoxia-inducible factor 1 (HIF-1) and is influenced by AMP-activated protein kinase (AMPK) signaling. Its activity directly increases glucose-6-phosphate levels, feeding into both glycolysis and the pentose phosphate pathway. ADPGK functionally interacts with glycolytic enzyme complexes, including hexokinase family proteins, and its role is intimately connected to the cellular energy status governed by the ADP/ATP ratio.
In the HT29 colorectal cancer context, ADPGK knockout cells provide a highly relevant model for dissecting metabolic reprogramming mechanisms that support tumor survival. Colorectal tumors often encounter hypoxic regions, and ADPGK’s ability to maintain glycolytic flux under low oxygen tension is thought to promote cancer cell viability. Disrupting ADPGK in HT29 cells allows researchers to elucidate how cancer cells adapt their energy metabolism through ADP-dependent glycolysis, and how this adaptation interfaces with other pathways such as HIF-1??-mediated transcription. This model is therefore significant for investigating the intersection of hypoxia response and metabolic reprogramming in colorectal adenocarcinoma.
Researchers can employ this knockout model in a wide range of experimental settings. Typical applications include investigating the role of ADPGK in colorectal cancer metabolism, studying hypoxia-induced metabolic reprogramming, and elucidating ADP-dependent glycolysis mechanisms. The cells are well-suited for metabolic flux analyses measuring glucose uptake and lactate production, proliferation and apoptosis assays under normoxia and hypoxia, and drug sensitivity screening with glycolysis inhibitors. Molecular assays such as Western blotting for ADPGK and HIF-1??, RT-qPCR for metabolic gene expression, and immunofluorescence for protein localization are also compatible. For more information, please contact Ascent Research.
