The AIFM2 Knockout MCF-7 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the MCF-7 human breast adenocarcinoma cell line, designed to disrupt the AIFM2 gene.
MCF-7 is an estrogen receptor-positive (ER+), progesterone receptor-positive (PR+), and HER2-low epithelial cell line originally established from the pleural effusion of a patient with metastatic mammary adenocarcinoma. This adherent, hormone-responsive model is widely used in breast cancer research, particularly for studying estrogen signaling, endocrine therapy resistance, and tumor progression.
AIFM2 encodes a mitochondrial flavoprotein oxidoreductase that functions as a key mediator of caspase-independent apoptosis. Transcriptionally induced by p53 in response to DNA damage or oxidative stress, AIFM2 is released from mitochondria and translocates to the nucleus, where it promotes chromatin condensation and large-scale DNA fragmentation. The protein interacts with FAD, NADH, cyclophilin D, and DNA, and cooperates with AIFM1 and EndoG in executing cell death. Upstream regulators include p53, E2F1, and reactive oxygen species, positioning AIFM2 at the convergence of intrinsic apoptosis and mitochondrial dysfunction pathways. Additionally, AIFM2 is implicated in redox regulation and maintaining mitochondrial morphology.
In the context of MCF-7 cells, which harbor wild-type p53, AIFM2-mediated apoptosis represents a critical pathway linking genotoxic stress to cell fate. Disruption of AIFM2 in this ER+ breast cancer model allows researchers to dissect the contribution of caspase-independent death mechanisms to chemotherapeutic responses, particularly to agents such as doxorubicin and etoposide. This model is instrumental for investigating chemoresistance and for screening AIFM2-dependent drug sensitivities.
Typical applications include the dissection of caspase-independent apoptotic signaling, functional studies of AIFM2 in mitochondrial biology, and elucidation of p53-mediated cell death pathways. Researchers can assess cell viability via MTT or CellTiter-Glo, measure apoptosis by Annexin V/PI flow cytometry or TUNEL assay, and monitor nuclear translocation through subcellular fractionation and immunofluorescence. Western blotting for AIFM2, cleaved caspase-3, and p53, along with mitochondrial membrane potential measurement using JC-1, enables comprehensive characterization. Drug sensitivity profiling and colony formation assays further support the study of chemoresistance mechanisms. For additional details or custom requirements, please contact Ascent Research.
