BCAT2 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population in which the BCAT2 gene has been disrupted within the HAP1 near-haploid human cell line. This knockout model abolishes the expression of mitochondrial branched-chain amino acid aminotransferase, enabling researchers to investigate the role of BCAT2 in BCAA catabolism and its downstream metabolic effects.
HAP1 cells are a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia cell line. Their haploid genetic architecture makes them an ideal host for functional genomics, as knockout of single-copy genes can be achieved with high efficiency. This system is extensively used for genetic screens and mechanistic studies due to the simplified allelic landscape, which facilitates unambiguous genotype-phenotype correlations.
BCAT2 encodes a mitochondrial branched-chain amino acid aminotransferase that catalyzes reversible transamination of leucine, isoleucine, and valine to their corresponding ??-keto acids, utilizing ??-ketoglutarate to generate glutamate. This reaction initiates BCAA degradation and connects amino acid catabolism to the TCA cycle. BCAT2 is transcriptionally regulated by PPAR?? and PGC-1??, requires pyridoxal phosphate as a cofactor, and produces ??-ketoisocaproate, ??-keto-??-methylvalerate, and ??-ketoisovalerate, which are then processed by the branched-chain ??-keto acid dehydrogenase complex into acetyl-CoA and succinyl-CoA.
The HAP1 background provides a streamlined genetic platform to study BCAT2 function with minimal interference from redundant alleles. Loss of BCAT2 in these cells perturbs BCAA catabolism, leading to alterations in cellular levels of branched-chain amino acids and their ??-keto acid derivatives, as well as reduced glutamate production and impaired TCA cycle anaplerosis. This knockout model is therefore valuable for dissecting metabolic adaptations in cancer cells, where BCAA metabolism is frequently reprogrammed, and for investigating inborn errors of BCAA metabolism such as hypervalinemia and hyperleucine-isoleucinemia.
Researchers can employ BCAT2 Knockout HAP1 Polyclonal Cells in a wide range of experimental applications, including Western blotting and RT-qPCR to confirm BCAT2 ablation, quantification of intracellular BCAA and ??-keto acid pools, Seahorse metabolic flux analysis to assess mitochondrial function, and isotopic tracing to map BCAA catabolic fates. The model also supports cell proliferation assays and high-throughput screening for BCAT2 inhibitors. For further information or to request this product, please contact Ascent Research.
