The BAX Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to eliminate BAX protein expression in the HCT 116 cell line. This polyclonal pool contains a heterogeneous mix of edited cells, each carrying independent loss-of-function alleles at the BAX locus, which averts clonal artifacts and provides a robust model for studying apoptosis resistance, chemoresistance, and non-apoptotic cell death mechanisms.
The HCT 116 cell line originates from a human colorectal carcinoma of a male patient and exhibits epithelial morphology with a near-diploid karyotype. It is characterized by an activating KRAS G13D mutation and high microsatellite instability (MSI-H) due to defective mismatch repair, and retains functional p53 signaling. These features make HCT 116 a powerful model for dissecting oncogenic pathways, DNA damage responses, and apoptotic regulation in a clinically relevant colorectal cancer context.
BAX functions as a critical pro-apoptotic effector of the BCL-2 family in the intrinsic mitochondrial apoptosis pathway. Under stress conditions such as DNA damage or growth factor withdrawal, TP53 transcriptionally upregulates BAX, and BH3-only proteins including BIM, BID, and PUMA trigger its conformational activation. Activated BAX oligomerizes at the mitochondrial outer membrane, where it interacts with VDAC and disrupts membrane integrity, leading to mitochondrial outer membrane permeabilization (MOMP). This event releases cytochrome c into the cytoplasm, which binds APAF1 to form the apoptosome, subsequently activating initiator caspase-9 and executioner caspases-3 and -7. BAX activity is tightly regulated by anti-apoptotic BCL-2 family members such as BCL-2, BCL-xL, and MCL-1 that sequester BAX, while 14-3-3 proteins retain it in an inactive cytosolic form.
By disrupting BAX in HCT 116 cells, this knockout model ablates the intrinsic apoptotic response, enabling the study of apoptosis-independent cell death modalities, including necroptosis and ferroptosis, and the assessment of chemoresistance mechanisms. The MSI-H and KRAS-mutant background of HCT 116 provides a relevant platform for investigating colorectal cancer biology, particularly for evaluating targeted agents and understanding how BAX loss impacts sensitivity to standard chemotherapies and novel pro-death compounds.
Key research applications include p53 pathway interrogation, compound screening for BAX-independent cell death inducers, and mitochondrial dysfunction analysis. Representative experimental techniques include Western blotting for BAX and cytochrome c release, annexin V/PI flow cytometry for apoptosis, caspase-3/7 activity measurements, JC-1 assays for mitochondrial membrane potential, and colony formation and IC50 profiling for long-term survival and drug sensitivity. For additional product details or to place an order, please contact Ascent Research.
