BCKDK Knockout AGS Cell Line

The BCKDK Knockout AGS Cell Line is a CRISPR/Cas9-edited knockout cell line providing targeted disruption of the BCKDK gene in a human gastric adenocarcinoma background. This loss-of-function model is designed to facilitate precise investigation of branched-chain amino acid (BCAA) catabolism and its regulatory roles in cellular metabolism and growth, offering a controlled experimental system without reliance on transient suppression methods.

AGS cells, derived from a human gastric adenocarcinoma, are widely used as an in vitro model for gastric cancer research. This adherent epithelial cell line retains key characteristics of gastric tumor biology, including sensitivity to metabolic and nutrient-sensing signals that are frequently dysregulated in gastric malignancies. The AGS background is particularly relevant for examining how tumor cells adapt their metabolic programs to sustain proliferation and survival.

BCKDK encodes the branched-chain alpha-ketoacid dehydrogenase kinase, which phosphorylates the BCKDHA subunit of the BCKDH complex, leading to inactivation of this rate-limiting enzyme in BCAA catabolism. Under nutrient-sufficient conditions, BCKDK is activated downstream of insulin signaling and mTORC1, reducing BCAA oxidation and promoting BCAA accumulation. Elevated BCAAs subsequently reinforce mTORC1 activation through S6K1 and S6 phosphorylation, establishing a feed-forward loop critical for coordinating amino acid availability with anabolic growth. BCKDK directly interacts with BCKDH complex components (BCKDHA, DBT, DLD) and is regulated by branched-chain alpha-ketoacids, positioning it as a central node connecting nutrient status to cellular energetics.

In gastric adenocarcinoma, aberrant BCAA metabolism and mTORC1 hyperactivation are linked to tumor progression and metabolic reprogramming. Disruption of BCKDK in AGS cells allows researchers to dissect the contribution of BCKDH complex regulation to gastric cancer cell metabolism, potentially revealing dependencies on BCAA-driven mTORC1 signaling for proliferation and survival. This model is valuable for testing whether BCKDK-mediated suppression of BCAA catabolism supports the anabolic demands of gastric cancer cells.

This knockout cell line supports diverse applications including western blot analysis of phospho-BCKDHA, phospho-S6K, and phospho-S6 to monitor mTORC1 activity, LC-MS-based BCAA quantification, and BCKDH enzymatic activity assays. It is also suitable for cell proliferation assays, metabolic vulnerability screening, and untargeted metabolomics profiling to identify pathway-specific alterations. Researchers can employ this model to study the interplay between nutrient availability, BCKDK action, and growth signaling in gastric cancer contexts. For further information, please contact Ascent Research.