The HK2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T human embryonic kidney epithelial cells. This product features targeted disruption of the HK2 gene, which encodes hexokinase 2, the enzyme catalyzing the first committed step of glycolysis. As a polyclonal mixture, it provides a heterogeneous loss-of-function model without clonal bias, suitable for robust metabolic and signaling studies.
The HEK293T host cell line stably expresses the SV40 large T antigen, enabling episomal replication of plasmids with an SV40 origin of replication. This feature enhances transient transfection efficiency and protein expression, making HEK293T a widely used platform for recombinant protein production and viral packaging. Derived from human embryonic kidney, these epithelial cells exhibit rapid growth and serve as an optimal background for generating knockout models to study pathways involved in metabolism, apoptosis, and oncogenic signaling.
HK2 phosphorylates glucose to glucose-6-phosphate in the initial step of glycolysis and is a pivotal node connecting metabolic flux to cell survival. Its expression is transcriptionally activated by c-MYC and HIF-1?? downstream of the PI3K/AKT/mTOR and AMPK pathways, while p53 represses HK2 transcription. At the mitochondrial outer membrane, HK2 interacts with VDAC1 to regulate mitochondrial permeability and is modulated by GSK3?? and AKT. This interaction links glycolytic output (pyruvate, lactate) to mTORC1 activity and cell proliferation signals, integrating nutrient availability with growth. Representative pathway components include PFK, PKM2, and LDH.
In HEK293T cells, HK2 knockout disrupts glycolytic flux, causing a metabolic shift from glycolysis to oxidative phosphorylation and a decrease in lactate production. Loss of the HK2-VDAC1 interaction sensitizes cells to apoptosis by altering mitochondrial membrane permeability, facilitating cytochrome c release, and reducing the anti-apoptotic protection normally conferred by mitochondrially bound HK2. This polyclonal knockout model thus captures the dual role of HK2 in metabolism and apoptosis, offering a powerful tool to dissect the Warburg effect and metabolic vulnerabilities in cancer cells.
Researchers can employ this model for cancer metabolism studies, glycolysis inhibition research, apoptosis pathway analysis, and drug target validation for metabolic reprogramming. Functional assessment may include Western blotting and RT-qPCR for HK2 expression, glucose uptake and lactate production assays, Seahorse metabolic flux analysis, annexin V apoptosis assays, mitochondrial membrane potential measurements, and co-immunoprecipitation of VDAC1. These applications support detailed bioenergetics profiling and investigation of therapeutic strategies targeting HK2. For further assistance, please contact Ascent Research.
