The AIFM2 Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the AIFM2 gene in the HCT 116 human colorectal carcinoma cell line. This product provides a heterogeneous pool of edited cells, offering a loss-of-function model to investigate AIFM2-mediated cell death pathways without the constraints of monoclonal selection. The polyclonal format captures the diversity of CRISPR-induced gene disruptions, enabling robust and reproducible functional studies in a genetically controlled background.
The HCT 116 cell line is a well-characterized model of human colorectal carcinoma, originating from a colonic epithelial tumor. It harbors a KRAS G13D activating mutation and retains wild-type TP53, while exhibiting MLH1 deficiency and high-level microsatellite instability (MSI-H). This genetic profile recapitulates key features of sporadic MSI-H colorectal cancers, where mismatch repair defects drive mutational burden and altered responses to DNA-damaging agents. The colonic epithelial origin makes HCT 116 a physiologically relevant platform for studying colorectal tumor biology, signal transduction, and therapeutic resistance mechanisms.
AIFM2 encodes a mitochondrial pro-apoptotic protein that mediates caspase-independent cell death via nuclear translocation upon apoptotic stimulation, directly inducing chromatin condensation and DNA fragmentation. In ferroptosis, AIFM2 facilitates iron-dependent lipid peroxidation, driving oxidative cell death. Mechanistically, AIFM2 functions downstream of TP53 and oxidative stress, with its activity modulated by chemotherapeutic agents and DNA damage. It physically interacts with BAX and BCL-2, influences cytochrome c release, and associates with COX4 in the intermembrane space. The ferroptosis regulators GPX4 and ACSL4 converge with AIFM2-mediated lipid peroxidation, positioning it at the intersection of apoptotic and ferroptotic pathways.
In the HCT 116 background, AIFM2 knockout provides a unique tool to dissect p53-dependent and -independent cell death mechanisms, given the cell line’s wild-type TP53 status and defective mismatch repair. The loss of AIFM2 may alter sensitivity to genotoxic chemotherapies, oxidative stressors, and ferroptosis inducers such as erastin or RSL3. This model enables researchers to examine how mitochondrial apoptosis and ferroptosis pathways are coordinated, and to identify synthetic lethal interactions that arise from combined KRAS mutation and AIFM2 deficiency, with implications for colorectal cancer therapeutic strategies.
This polyclonal knockout product is suited for a broad range of experimental applications, including apoptosis research via Annexin V flow cytometry, TUNEL assays, and mitochondrial membrane potential measurements; ferroptosis studies using lipid ROS detection and GPX4/ACSL4 expression analysis; and p53 signaling investigations through quantitative PCR and RNA-seq. It facilitates cell viability assays under chemotherapeutic challenge, Western blotting for AIFM2 and downstream effectors, and immunofluorescence to monitor AIFM2 subcellular localization. Researchers can employ this model to explore drug resistance mechanisms, validate target engagement, and conduct genetic interaction screens. For additional product details or technical support, please contact Ascent Research.
