The BAX Knockout DLD-1 Polyclonal Cells product provides a polyclonal population of DLD-1 cells with CRISPR/Cas9-mediated disruption of the BAX gene. This knockout eliminates BAX, a pro-apoptotic Bcl-2 family protein required for mitochondrial outer membrane permeabilization and cytochrome c release, central to intrinsic apoptosis. The polyclonal nature captures diverse editing outcomes, enabling pooled screens and pathway analyses without isolation artifacts.
DLD-1 (ATCC CCL-221) is an epithelial colorectal adenocarcinoma cell line from a male patient, widely used as a colorectal cancer model. These cells retain key oncogenic pathways, including aberrant Wnt/??-catenin and PI3K/AKT signaling, providing a clinically relevant system for apoptosis and drug resistance studies.
BAX functions downstream of diverse stress signals and is activated by BH3-only proteins such as BIM, PUMA, and tBID, which neutralize anti-apoptotic BCL2, BCL-xL, and MCL1. Upon activation, BAX translocates to mitochondria, oligomerizes with BAK1, and forms pores that release cytochrome c and SMAC/DIABLO. Cytochrome c binds Apaf-1 to activate caspase-9, which cleaves caspase-3, committing the cell to apoptosis. p53 transcriptionally upregulates BAX, while kinases like JNK, AKT, and ERK modulate its activity. Thus, BAX serves as an essential gatekeeper of mitochondrial apoptosis.
In DLD-1 cells, BAX knockout abolishes mitochondrial apoptosis, enabling dissection of BAX-dependent versus BAX-independent death pathways. This model is particularly relevant for colorectal cancer, where therapeutic resistance often involves suppression of apoptosis. It allows identification of bypass mechanisms and testing of BAX-targeted therapies, as well as evaluation of alternative death modalities such as necroptosis.
Applications span apoptosis signaling studies, anticancer drug resistance profiling, and high-throughput screening for pro-apoptotic compounds. Representative assays include Western blotting for BAX, cytochrome c, and cleaved caspase-3; flow cytometry with annexin V-FITC/PI; JC-1 mitochondrial membrane potential measurement; cell viability (MTT) and caspase-3/7 Glo assays; colony formation; and immunofluorescence for BAX translocation. Co-immunoprecipitation can probe BAX interactions with BAK1 or BCL2. This knockout pool also enables functional genomics screens in colorectal cancer. For technical inquiries and ordering information, please contact Ascent Research.
