The BCAT2 Knockout HCT 116 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HCT 116 cells with targeted disruption of the BCAT2 gene. This heterogeneous knockout pool enables loss-of-function studies without clonal selection, providing a flexible system for investigating branched-chain amino acid (BCAA) metabolism in a colorectal carcinoma background.
The HCT 116 host cell line is a human colorectal carcinoma epithelial model featuring oncogenic mutations in KRAS (G13D), PIK3CA, and ??-catenin, along with microsatellite instability (MSI). These cells display robust epithelial adherence and proliferation, making them a relevant platform for studying cancer cell biology and metabolic reprogramming.
BCAT2 encodes a mitochondrial branched-chain amino acid aminotransferase that catalyzes the transamination of leucine, isoleucine, and valine to ??-ketoglutarate, producing glutamate and branched-chain ??-keto acids. This reaction links BCAA catabolism to the TCA cycle and is regulated by upstream factors MYC and HIF1??, as well as by leucine availability and mTORC1 signaling. BCAT2 activity influences mTORC1 activation and interacts with the branched-chain ??-keto acid dehydrogenase (BCKDH) complex, positioning it as a key node in nutrient sensing and amino acid homeostasis.
In the HCT 116 background, BCAT2 disruption impairs BCAA degradation, likely altering intracellular BCAA, ??-ketoglutarate, and glutamate pools. This metabolic perturbation may attenuate mTORC1 signaling, disrupt TCA cycle anaplerosis, and compromise amino acid homeostasis. Given the MSI-H and KRAS-mutant status, BCAT2 loss could reveal metabolic vulnerabilities and synthetic lethal interactions pertinent to colorectal cancer.
Researchers can utilize this knockout product to examine BCAA metabolism in colon cancer, mTORC1 nutrient sensing, and amino acid addiction. Validation assays include Western blot, RT-qPCR, and Sanger sequencing; functional studies encompass metabolomics, phospho-signaling analysis (p-S6K, p-4E-BP1), proliferation, apoptosis, and amino acid uptake assays. For further information, please contact Ascent Research.
