The GLS Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the GLS gene in HEK293T cells. This model disrupts the expression of glutaminase, the enzyme responsible for catalyzing the hydrolysis of glutamine to glutamate, thereby impairing a central metabolic node. The polyclonal nature of the population ensures representation of diverse editing events, providing a heterogeneous loss-of-function model suitable for studying the collective effects of GLS disruption without the constraints of clonal variation. As a CRISPR/Cas9-mediated gene disruption tool, these cells enable investigation of glutamine metabolism in a human kidney epithelial background.
The host cell line, HEK293T, is an immortalized human embryonic kidney epithelial cell line stably expressing the SV40 large T antigen. This genetic modification confers high transfection efficiency and robust protein expression, making HEK293T a preferred system for exogenous DNA uptake and recombinant protein production. Derived from the parental HEK293 line, these cells exhibit adherent growth and maintain characteristics of kidney epithelium, including the expression of relevant transporters and metabolic enzymes. Their rapid proliferation and amenability to genetic manipulation establish a consistent background for metabolic and signaling studies.
GLS encodes glutaminase, which catalyzes the first step in glutamine utilization by converting glutamine to glutamate, a precursor for the tricarboxylic acid (TCA) cycle intermediate ??-ketoglutarate. This reaction is transcriptionally activated by the oncogene MYC and regulated upstream by mTORC1 and the tumor suppressor p53, placing GLS at the intersection of growth factor signaling and nutrient sensing. Downstream, glutamate contributes to the synthesis of glutathione via SIRT5-mediated modifications and serves as a building block for non-essential amino acids. GLS also interacts with protein phosphatase 2A (PP2A), linking metabolic flux to signaling networks. Representative pathway components include glutamine, glutamate, ??-ketoglutarate, and TCA cycle intermediates that feed nucleotide and lipid biosynthesis.
In the HEK293T background, knockout of GLS profoundly alters cellular metabolism by eliminating the primary source of glutamine-derived ??-ketoglutarate, thereby disrupting anaplerotic replenishment of the TCA cycle. This perturbation reduces the availability of metabolic intermediates required for ATP production, amino acid synthesis, and glutathione-dependent redox balance. Consequently, GLS-null HEK293T cells exhibit impaired proliferation and metabolic reprogramming, making them a powerful model for dissecting the role of glutamine metabolism in transformed cells. The system is particularly relevant given that HEK293T cells share features with cancer cells in their reliance on glutamine for growth and survival, offering a platform to investigate the consequences of GLS loss in a controlled, experimentally tractable context.
These polyclonal GLS knockout cells are ideally suited for a variety of research applications, including cancer metabolism studies, glutamine dependency assays, and drug target validation. They enable detailed metabolic flux analyses using glutaminase activity measurements, metabolomics, and TCA cycle tracing, as well as functional assays such as Seahorse metabolic profiling, Western blotting for downstream targets, and cell proliferation assays. Researchers can dissect mTORC1 signaling, glutathione synthesis, and ??-ketoglutarate production in a model that uncouples glutamine availability from glutamate generation. For additional technical specifications, protocols, or pricing, please contact Ascent Research.
