The AIFM2 knockout HT29 polyclonal cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HT29 human colorectal adenocarcinoma line, providing targeted disruption of the AIFM2 gene. This knockout tool enables precise investigation of AIFM2-mediated signaling pathways in a well-characterized intestinal epithelial cancer model, supporting advanced studies in ferroptosis, caspase-independent apoptosis, and redox biology.
HT29 cells, isolated from a 44-year-old female with colorectal adenocarcinoma, serve as a canonical model for colorectal cancer research, intestinal epithelial barrier function, and drug metabolism studies. Their epithelial origin and robust growth characteristics make them particularly suitable for probing pathways governing apoptosis resistance and mitochondrial redox homeostasis in colon cancer, offering a physiologically relevant context for genetic perturbation.
AIFM2 encodes a mitochondrial flavoprotein oxidoreductase that functions as a key mediator of caspase-independent apoptosis through nuclear translocation and chromatin degradation, while also promoting ferroptosis by enhancing lipid peroxidation via its oxidoreductase activity. The protein is transcriptionally activated by p53/p73 family members under oxidative or genotoxic stress, and operates downstream of DNA damage and hypoxia signaling. AIFM2 interacts with flavoprotein redox partners, cyclophilin A, and its homolog AIFM1, and engages ferroptotic machinery including GPX4, ACSL4, and iron-dependent lipid peroxide accumulation. In the intrinsic apoptosis pathway, it acts in parallel with BAX/BAK, cytochrome c, and APAF1, while within the p53 network it aligns with downstream effectors p21 and PUMA.
In the HT29 adenocarcinoma context, AIFM2 disruption enables dissection of its dual role in ferroptosis and caspase-independent apoptosis, pathways often dysregulated in colorectal cancer. This model is uniquely suited to exploring how mitochondrial redox regulation and lipid peroxidation drive cell death decisions in intestinal epithelial cells, and to evaluating therapeutic vulnerabilities in apoptosis-resistant cancers where AIFM2 may serve as a critical node linking oxidative stress to cell fate.
Typical research applications include mechanistic studies of ferroptosis induction using erastin or RSL3, analysis of caspase-independent cell death, and validation of AIFM2 as a drug target in colorectal cancer. Compatible assays include Western blot for AIFM2 and GPX4, RT-qPCR, Annexin V/PI apoptosis assay, lipid peroxidation measurement with C11-BODIPY, GPX4 activity assays, mitochondrial membrane potential analysis, and comet assay for DNA damage. For further information on this knockout tool, please contact Ascent Research.
