This product is a CRISPR/Cas9-edited polyclonal knockout HT29 cell population, derived from a human colorectal adenocarcinoma line, with targeted disruption of the BCAT2 gene locus. The knockout abolishes BCAT2 protein expression across the polyclonal pool, providing a loss-of-function model for studying branched-chain amino acid transaminase 2 in colon cancer biology. Its polyclonal nature maintains cellular heterogeneity while ensuring consistent target-gene disruption, making it a versatile tool for functional studies.
HT29 cells are a well-established human colorectal adenocarcinoma line, originally isolated from a primary tumor of a 44-year-old female patient. They display adherent epithelial morphology and are extensively used as a model for colon cancer, including investigations of drug resistance, differentiation, and oncogenic signaling pathways. The cell line’s robust growth characteristics and genetic background make it an ideal host for CRISPR-mediated gene editing, enabling the creation of derivative knockout models that retain key features of the original tumor.
BCAT2 encodes a mitochondrial branched-chain amino acid aminotransferase that catalyzes the reversible transamination of leucine, isoleucine, and valine to their corresponding alpha-keto acids and glutamate. This reaction represents the initial and rate-limiting step in BCAA catabolism, directing carbon skeletons into the TCA cycle and de novo lipid synthesis. BCAT2 activity is intimately connected to mTORC1 signaling: its alpha-keto acid products promote mTORC1 activation via Rag GTPases, while BCAT2 transcription is regulated by oncogenic drivers MYC and HIF1A, as well as by nutrient availability. In turn, the BCAT2 metabolic axis collaborates with the BCKDH complex, glutamate dehydrogenase, and other aminotransferases to integrate amino acid degradation with cellular energy status and anabolic growth signals.
Within HT29 colorectal cancer cells, BCAT2-mediated BCAA catabolism is thought to meet the elevated metabolic demands of rapid proliferation, providing substrates for the TCA cycle and lipid biosynthesis, and sustaining mTORC1-driven growth programs. Disruption of BCAT2 through CRISPR/Cas9 gene editing is expected to impair these metabolic fluxes, potentially leading to reduced cell growth, altered stress responses, and heightened sensitivity to nutrient deprivation. Consequently, this knockout model enables detailed interrogation of how loss of BCAT2 reshapes metabolic reprogramming, redox homeostasis, and signaling networks in a colorectal cancer context, revealing therapeutic targets linked to BCAA metabolism.
This polyclonal knockout cell product supports diverse research applications, including metabolic flux analysis using isotope-labeled BCAAs, BCAT2 enzymatic activity assays, and western blotting for BCAT2 and phospho-S6K to evaluate mTORC1 pathway activity. Further experimental approaches encompass RT-qPCR profiling of BCAA metabolic enzymes, cell proliferation and colony formation assessments, apoptosis detection, and migration or invasion assays, enabling comprehensive functional studies of BCAT2 in colorectal cancer progression, metastasis, and drug resistance. For technical details or to request pricing, please contact Ascent Research.
