ADPGK Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout cell population generated from the HCT 116 human colorectal carcinoma cell line, designed to disrupt the ADPGK gene (ADP-dependent glucokinase). This loss-of-function model allows researchers to investigate the role of an alternative glucose phosphorylation pathway in cancer metabolism. The polyclonal format provides a heterogeneous knockout population directly edited by CRISPR/Cas9 without clonal isolation, reflecting a pool of genetic variants that collectively ablate ADPGK function. Cell populations are validated for reduced target protein expression and are suitable for a range of downstream functional assays.
The HCT 116 host cell line is a well-characterized model of colorectal carcinoma, derived from a male patient, exhibiting a near-diploid karyotype and harboring a homozygous mutation in the MLH1 gene, resulting in microsatellite instability (MSI-H). This genetic background makes HCT 116 cells particularly relevant for studying DNA mismatch repair-deficient cancers. Widely used in tumor biology, HCT 116 cells support robust proliferation and have been extensively employed in drug screening, oncogenic signaling studies, and metabolic profiling. Their epithelial origin and colorectal cancer context provide a physiologically relevant platform for examining metabolic reprogramming.
ADPGK encodes an ADP-dependent glucokinase that catalyzes the phosphorylation of glucose to glucose-6-phosphate using ADP as the phosphate donor, representing a non-canonical entry point into glycolysis and the pentose phosphate pathway (PPP). ADPGK activity is transcriptionally regulated by factors such as HIF1A and MYC, and is modulated by energy-sensing AMPK signaling and the carbohydrate-responsive element-binding protein ChREBP. Downstream, ADPGK contributes to the production of glucose-6-phosphate, ribose-5-phosphate, NADPH, and lactate. The protein interacts with canonical pathway components including hexokinase (HK), phosphofructokinase (PFKL), pyruvate kinase (PKM), lactate dehydrogenase A (LDHA), glucose-6-phosphate dehydrogenase (G6PD), and transketolase (TKT). Disruption of ADPGK impairs flux through both glycolysis and the PPP, reducing biosynthetic precursors and compromising cellular redox balance.
In HCT 116 colorectal cancer cells, ADPGK knockout disrupts the alternative glycolytic route, highlighting the dependency of these cells on ADP-dependent glucose utilization under specific metabolic conditions. This model is instrumental for dissecting how cancer cells adapt glucose metabolism in the context of MLH1 deficiency and genomic instability. The knockout cells enable exploration of the interplay between conventional hexokinase-driven glycolysis and the ADPGK-mediated bypass, particularly relevant under hypoxic or nutrient-stressed tumor microenvironments where HIF1A and MYC are active. Consequently, this system serves as a valuable tool for validating metabolic vulnerabilities and potential therapeutic targets in colorectal cancer.
Typical research applications include metabolic flux analysis using Seahorse extracellular flux analyzers to measure ECAR and OCR, quantitative assessment of glucose consumption and lactate secretion, and NADPH/NADP+ ratio measurements. Standard validation assays such as Western blotting for ADPGK protein and RT-qPCR for ADPGK mRNA confirm target disruption. The polyclonal knockout cells are suitable for proliferation assays (MTT, BrdU), clonogenic survival, apoptosis detection via Annexin V staining, and flow cytometric cell cycle analysis. This product supports studies in cancer metabolism, alternative glycolytic pathway investigation, drug target validation, and metabolic reprogramming in colorectal carcinoma. For additional information or technical support, please contact Ascent Research.
